Hypoimmunogenic CD22-targeted car t cells for the treatment of relapsed and / or refractory b cell lymphomas

Abstract

The present disclosure provides methods of treating a subject having or suspected of having a B cell malignancy or a B cell mediated disorder by administering hypoimmune allogenic CD22-directed CAR T cells. Also disclosed are pharmaceutical compositions comprising hypoimmune allogenic CD22-directed CAR T cells, for use in treating a subject having or suspected of having a B cell malignancy or a B cell mediated disorder.

Description

Attorney Docket No. 2017428-0684 METHODS AND COMPOSITIONS FOR TREATING B-CELL MALIGNANCIES CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of United States Provisional Application No. 63 / 595,295, filed November 1, 2023 and United States Provisional Application No. 63 / 659,201, filed June 12, 2024; the contents of which are incorporated herein by reference in their entirety. BACKGROUND

[0002] B cell lymphomas encompass a group of lymphoma subtypes categorized within 2 broad categories: non-Hodgkin lymphoma (NHL) and Hodgkin lymphoma (HL), with NHLs being the majority of diagnosed cases. The therapeutic landscape varies for each distinct tumor subtype, but generally there are limited effective therapies in relapsed and / or refractory (r / r) settings across B cell lymphomas.

[0003] Despite recent advancement, NHL remains an unmet medical need. In 2020, there were 123,000 new cases of NHL reported with 50,000 attributed deaths in Europe. NHL is a heterogeneous group of cancers originating in B lymphocytes, T lymphocytes, or natural killer (NK) cells. In the US, B cell lymphomas represent approximately 85% of all NHL cases.

[0004] There is an urgent need to develop therapies for such patients. SUMMARY

[0005] Off-the-shelf CAR-T cells and other therapeutic cells can offer advantages over autologous cell- based strategies, including ease of manufacturing, quality control, and avoidance of malignant contamination and T cell dysfunction. However, the vigorous host-versus-graft immune response against histoincompatible T cells prevents expansion and persistence of allogeneic CAR-T cells and mitigates the efficacy of this approach.

[0006] There is substantial evidence in both animal models and human patients that hypoimmunogenic cell transplantation is a scientifically feasible and clinically promising approach to the treatment of numerous disorders, conditions, and diseases.

[0007] There remains a need for novel approaches, compositions and methods for producing cell- based therapies that avoid detection by the recipient’s immune system.

[0008] The present disclosure provides methods of treating a subject having or suspected of having a B cell malignancy or a B cell mediated disorder. In some embodiments, the method comprises: administering to the subject a composition comprising a population of engineered hypoimmunogenic T cells. In some embodiments, the hypoimmunogenic T cells comprise one or more modifications. In some embodiments, the one or more modifications inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and / or (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules. In some Page 1 of 173 12403779v1Attorney Docket No. 2017428-0684 embodiments, the one or more modifications increase expression of CD47 encoded by a first exogenous polynucleotide. In some embodiments, increased expression of CD47 encoded by a first exogenous polynucleotide is relative to a control or wild-type T cell that does not comprise the modification. In some embodiments, the one or more modifications express a CD22-directed chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.

[0009] In some embodiments, one or more modifications reduce expression of one or more MHC class I molecules, one or more MHC class II molecules, and / or one or more TCR molecules in the hypoimmunogenic T cells, relative to control or wild-type T cells. In some embodiments, expression of one or more MHC class I molecules, one or more MHC class II molecules, and one or more TCR molecules is reduced in the hypoimmunogenic T cells relative to a control or wild-type T cell. In some embodiments, a control or wild-type T cells do not comprise the modifications. In some embodiments, one or more modifications reduce cell surface expression of the one or more MHC class I molecules relative to a control or wild-type T cell. In some embodiments, one or more modifications reduce cell surface trafficking of one or more MHC class I molecules relative to a control or wild-type T cell. In some embodiments, one or more modifications reduce a function of one or more MHC class I molecules. In some embodiments, one or more MHC class I molecules is one or more human leukocyte antigen (HLA) class I molecules. In some embodiments, one or more HLA class I molecules is HLA-A, HLA-B, HLA-C, or a combination thereof.

[0010] In some embodiments, one or more molecules that regulate cell surface expression of one or more MHC class I molecules, regulate cell surface trafficking of the one or more MHC class I molecules, and / or reduce a function of one or more MHC class I molecules are beta-2 microglobulin (B2M). In some embodiments, one or more modifications comprise a modification that inactivates or disrupts one or more alleles of B2M. In some embodiments, a modification that inactivates or disrupts one or more alleles of B2M reduces mRNA expression of the B2M gene. In some embodiments, a modification that inactivates or disrupts one or more alleles of B2M reduces protein expression of B2M. In some embodiments, a modification that inactivates or disrupts one or more alleles of B2M comprises: inactivation or disruption of one allele of the B2M gene; inactivation or disruption of both alleles of the B2M gene; or inactivation or disruption of all B2M coding alleles in a hypoimmunogenic T cell. In some embodiments, a modification that inactivates or disrupts one or more alleles of B2M comprises an indel in the B2M gene. In some embodiments, a modification that inactivates or disrupts one or more alleles of B2M comprises a frameshift mutation or a deletion of a contiguous stretch of genomic DNA of the B2M gene.

[0011] In some embodiments, one or more modifications reduce cell surface expression of the one or more MHC class II molecules relative to a control or wild-type cell. In some embodiments, one or more modifications reduce cell surface trafficking of the one or more MHC class II molecules relative to a control or wild-type cell. In some embodiments, one or more modifications reduce a function of one or more MHC class II molecules. In some embodiments, function of the one more MHC class II molecules is antigen presentation. In some embodiments, one or more MHC class II molecules is one or more human leukocyte antigen (HLA) class II molecules. In some embodiments, one or more HLA class II molecules is HLA-DP, HLA-DQ, HLA-DR, or a combination thereof.

[0012] In some embodiments, one or more molecules that regulate expression of the one or more MHC class II molecules are Class II Major Histocompatibility Complex Transactivator (CIITA). In some embodiments, one or more modifications comprise a modification that inactivates or disrupts one or more alleles Page 2 of 173 12403779v1Attorney Docket No. 2017428-0684 of CIITA. In some embodiments, a modification that inactivates or disrupts one or more alleles of CIITA reduces mRNA expression of the CIITA gene. In some embodiments, a modification that inactivates or disrupts one or more alleles of CIITA reduces protein expression of CIITA. In some embodiments, a modification that inactivates or disrupts one or more alleles of CIITA comprises: inactivation or disruption of one allele of the CIITA gene; inactivation or disruption of both alleles of the CIITA gene; or inactivation or disruption of all CIITA coding alleles in the hypoimmunogenic T cell. In some embodiments, a modification that inactivates or disrupts one or more alleles of CIITA comprises an indel in a CIITA gene. In some embodiments, a modification that inactivates or disrupts one or more alleles of CIITA comprises a frameshift mutation or a deletion of a contiguous stretch of genomic DNA of a CIITA gene.

[0013] In some embodiments, expression of HLA-A, HLA-B, HLA-C, HLA-DP, HLA-DQ, and HLA-DR are reduced in a hypoimmunogenic T cell. In some embodiments, one or more modifications reduce cell surface protein expression of one or more TCR molecules relative to the control or wild-type cell. In some embodiments, one or more modifications reduce cell surface trafficking of the one or more TCR molecules relative to a control or wild-type cell. In some embodiments, one or more modifications reduce a function of one or more TCR molecules. In some embodiments, a function of one or more TCR molecules is antigen recognition. In some embodiments, one or more TCR molecules is T cell receptor alpha constant (TRAC), T cell receptor beta constant (TRBC), or a combination thereof. In some embodiments, one or more TCR molecules comprise TRAC. In some embodiments, one or more modifications comprise a modification that inactivates or disrupts one or more alleles of TRAC. In some embodiments, a modification that inactivates or disrupts one or more alleles of TRAC reduces mRNA expression of a TRAC gene. In some embodiments, a modification that inactivates or disrupts one or more alleles of TRAC reduces protein expression of TRAC. In some embodiments, a modification that inactivates or disrupts one or more alleles of TRAC comprises: inactivation or disruption of one allele of the TRAC gene; inactivation or disruption of both alleles of the TRAC gene; or inactivation or disruption of all TRAC coding alleles in a hypoimmunogenic T cell. In some embodiments, a modification inactivation or disruption that inactivates or disrupts one or more alleles of TRAC comprises an indel in the TRAC gene. In some embodiments, inactivation or disruption comprises a frameshift mutation or a deletion of a contiguous stretch of genomic DNA of the TRAC gene.

[0014] In some embodiments, one or more modifications increase expression of one or more tolerogenic factors encoded by a third exogenous polynucleotide, wherein the increased expression of the one or more tolerogenic factors is relative to a control or wild-type T cell, and wherein the one or more tolerogenic factors comprise CD16, CD24, CD35, CD39, CD46, CD47, CD52, CD55, CD59, CD64, CD200, CCL22, CTLA4-Ig, C1 inhibitor, FASL, IDO1, HLA-C, HLA-E, HLA-E heavy chain, HLA-G, IL-10, IL-35, PD-L1, SERPINB9, CCL21, MFGE8, DUX4, B2M-HLA-E, CD27, IL-39, CD16 Fc Receptor, IL15-RF, H2-M3 (HLA-G), A20 / TNFAIP3, CR1, HLA-F, MANF, or a combination thereof.

[0015] The present disclosure provides methods of treating a subject having or suspected of having a toPage 3 of 173 12403779v1Attorney Docket No. 2017428-0684 relative to the control or wild-type T cells, and (c) expression of a CD22-directed chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.

[0016] In some embodiments, a composition is administered to the subject at a dosage of about 5x106cells to about 400x106cells. In some embodiments, a composition is administered to the subject at a dosage of about 40x106cells to about 250x106cells. In some embodiments, a composition is administered to the subject at a dosage of about 20x106cells to about 400x106cells. In some embodiments, a composition is administered to the subject at a dosage of at least about 30x106cells, at least about 40x106cells, at least about 60x106cells, at least about 120x106cells, at least about 150x106cells, at least about 200x106cells, or at least about 250x106cells. In some embodiments, a composition is administered to the subject at a dosage of no more than about 200x106cells. In some embodiments, a composition is administered to the subject at a dosage of no more than about 250x106cells. In some embodiments, a composition is administered to the subject at a dosage of no more than about 400x106cells. In some embodiments, a composition is administered to the subject at a dosage of about 5x106CAR+ cells to about 400x106CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of about 20x106CAR+ cells to about 400x106CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of about 40x106CAR+ cells to about 250x106CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of at least about 30x106CAR+ cells, at least about 60x106CAR+ cells, at least about 120x106CAR+ cells, or at least about 200x106CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of no more than about 200x106CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of no more than about 400x106CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of about 20x106CD22-directed CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of about 60x106CD22-directed CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of about 120x106CD22-directed CAR+ cells. In some embodiments, a composition is administered to the subject at a dosage of about 200x106CD22- directed CAR+ cells.

[0017] In some embodiments, a total volume of about 1.25mL to about 100mL of the composition is administered. In some embodiments, a total volume of about 5mL to about 100mL of the composition is administered. In some embodiments, a total volume of at least about 7.5mL, at least about 15mL, at least about 30mL, or at least about 50mL of the composition is administered. In some embodiments, a composition comprises a concentration of engineered hypoimmunogenic T cells of at least 1.25x106cells / mL. In some embodiments, a composition comprises a concentration of engineered hypoimmunogenic T cells of at least 4.0x106cells / mL. In some embodiments, a composition comprises a concentration of engineered hypoimmunogenic T cells of at least 1.2x106CAR+ cells / mL. In some embodiments, a composition comprises a concentration of engineered hypoimmunogenic T cells of at least 4.0x106CAR+ cells / mL.

[0018] In some embodiments, methods of the present disclosure provide administering a composition in at least one dose, at least two doses, at least three doses, at least four doses, at least five doses, at least six doses, at least seven doses, at least eight doses, at least ten doses, at least eleven doses, or at least twelve doses. In some embodiments, methods of the present disclosure provide administering the composition in a single dose. In some embodiments, methods of the present disclosure provide administering the composition in two doses. Page 4 of 173 12403779v1Attorney Docket No. 2017428-0684

[0019] In some embodiments, engineered hypoimmunogenic T cells comprise CD8+ and / or CD4+ T cells. In some embodiments, engineered hypoimmunogenic T cells comprise allogeneic T cells. In some embodiments, engineered hypoimmunogenic T cells comprise primary T cells. In some embodiments, engineered hypoimmunogenic T cells comprise stem cell-derived T cells.

[0020] In some embodiments, a CAR comprises an anti-CD22 antigen binding domain, a CD8α hinge domain, a CD8α transmembrane domain, a 4-1BB costimulatory domain, and a CD3ζ intracellular signaling a 37.

[0023] In some embodiments, a B cell malignancy is a lymphoma or a leukemia. In some embodiments, a B cell malignancy is selected from the group consisting of: Non-Hodgkin’s Lymphoma (NHL), Chronic Lymphocytic Leukemia (CLL), large B cell lymphoma (LBCL), transplant ineligible large B cell lymphoma (LBCL), diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), primary mediastinal B cell lymphoma (PMBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), marginal zone lymphoma (MZL), or small lymphocytic lymphoma (SLL). Page 5 of 173 12403779v1Attorney Docket No. 2017428-0684

[0024] In some embodiments, a subject is diagnosed to have NHL or CLL. In some embodiments, a subject is diagnosed to have NHL based on the World Health Organization 2016 criteria. In some embodiments, a subject is confirmed to have NHL based on histological analysis of a biopsy sample collected from the subject. In some embodiments, a subject has: a single lesion measuring greater than 7.5 cm in diameter; or a sum of the product of lymph node diameter is greater than 5000 mm3.

[0025] In some embodiments, a subject is diagnosed to have CLL based on the International Workshop on CLL (iwCLL) criteria. In some embodiments, a subject has: a lymph node greater than or equal to 10 cm in diameter; or a lymph node greater than or equal to 5 cm in diameter and an absolute lymphocyte count greater than or equal to 25 X 109lymphocytes / L.

[0026] In some embodiments, a subject is diagnosed as being a high risk CCL patient. In some embodiments, a method of the present disclosure further comprises: detecting one or more biomarkers, and identifying a subject as being a high risk CCL patient based on the one or more biomarkers. In some embodiments, one or more biomarkers are selected from the group consisting of: del(17p), mutated TP53, unmutated immunoglobulin heavy-chain variable (IGHV) gene, and complex karyotype.

[0027] In some embodiments, a subject previously received an alkylating agent, a purine nucleoside, chemoimmunotherapy, a Bruton’s tyrosine kinase (BTK) inhibitor, a PI3K inhibitor, a B cell lymphoma 2 (BCL2) inhibitor, a CD20 monoclonal antibody, a bispecific monoclonal antibody, or a combination thereof. In some embodiments, an alkylating agent comprises chlorambucil or cyclophosphamide. In some embodiments, a purine nucleoside is selected from the group consisting of: fludarabine, pentostatin, and cladribine. In some embodiments, a chemoimmunotherapy is selected from the group consisting of: FCR (fludarabine, cyclophosphamide, and rituximab), BR (bendamustine and rituximab), alemtuzumab, and lenalidomide. In some embodiments, a BTK inhibitor is selected from the group consisting of: ibrutinib, acalabrutinib, zanubrutinib, and pirtorutinib. In some embodiments, a PI3K inhibitor comprises idelalisib or duvelisi. In some embodiments, a BCL2 inhibitor comprises venetoclax. In some embodiments, a CD20 monoclonal antibody comprises obinutuzumab. In some embodiments, a bispecific monoclonal antibody is selected from the group consisting of Mosunetuzumab (CD20 x CD3 mAb), Glofitamab (CD20(2) x CD3), Odronextamab (CD20 x CD3 mAb), and Epcoritamab (CD20 x CD3 mAb).

[0028] In some embodiments, a subject is diagnosed to have CLL and is further administered a BTK inhibitor. In some embodiments, a BTK inhibitor is selected from the group consisting of: ibrutinib, acalabrutinib, zanubrutinib, and pirtorutinib.

[0029] In some embodiments, a subject is diagnosed to have CLL and is administered the composition comprising the population of engineered hypoimmunogenic T cells as a first-line therapy.

[0030] In some embodiments, a B cell malignancy or the B cell mediated disorder is relapsed or refractory following at least one therapeutic standard of care regimen. In some embodiments, a B cell malignancy or the B cell mediated disorder is relapsed or refractory following at least two therapeutic standard of care regimens. In some embodiments, a B cell malignancy or the B cell mediated disorder is relapsed or refractory following at least three therapeutic standard of care regimens. In some embodiments, a B cell malignancy or the B cell mediated disorder is relapsed or refractory following autologous stem cell transplant (ASCT). In some embodiments, a subject: did not respond to a first line therapy; had progressive B cell malignancy or B cell mediated disorder as a best response to a last administered therapeutic regimen; or had Page 6 of 173 12403779v1Attorney Docket No. 2017428-0684 stable disease as a best response to the last administered therapeutic regimen. In some embodiments, a subject: had disease progression or relapse within twelve months of ASCT; or the subject had no response to or relapsed following administration of a last line of therapy.

[0031] In some embodiments, a subject has adequate hepatic function, renal function, bone marrow function, cardiac function, and / or pulmonary function.

[0032] In some embodiments, a B cell malignancy is relapsed and / or refractory. In some embodiments, a B cell malignancy is relapsed and / or refractory NHL. In some embodiments, a B cell malignancy is relapsed and / or refractory CLL. In some embodiments, a B cell malignancy is relapsed and / or refractory after at least 2 regimens of therapy. In some embodiments, a B cell malignancy is relapsed and / or refractory after at least 3 regimens of therapy. In some embodiments, a B cell malignancy is relapsed and / or refractory after autologous stem cell transplant (ASCT). In some embodiments, a B cell malignancy is relapsed and / or refractory, wherein relapsed and / or refractory comprises one or more of: no response to first line therapy, progressive disease as best response to most recent therapy regimen, stable disease as best response to most recent therapy with duration no longer than 6 months from last dose of therapy, disease progression or relapse within 12 months of ASCT, no response to or relapse after last line of therapy post-ASCT.

[0033] In some embodiments, a subject has not received a prior CD22-directed therapy. In some embodiments, a subject has received a prior CD22-directed therapy. In some embodiments, a prior CD22- directed therapy is or comprises a CD22-directed CAR T cell treatment. In some embodiments, a prior CD22- directed therapy is or comprises a CD22-directed antibody treatment. In some embodiments, a prior CD22- directed therapy is or comprises a CD22-directed CAR natural killer (NK) cell treatment.

[0034] In some embodiments, a subject has received a prior bispecific antibody therapy. In some embodiments, a bispecific antibody therapy is selected from the group consisting of: Mosunetuzumab (CD20 x CD3 mAb), Glofitamab (CD20(2)x CD3), Odronextamab (CD20 x CD3 mAb), and Epcoritamab (CD20 x CD3 mAb).

[0035] In some embodiments, a subject does not have a history of primary central nervous system (CNS) lymphoma or CNS metastases. In some embodiments, absence of primary CNS lymphoma or CNS metastases is based on a brain magnetic resonance imaging (MRI) performed within 3 months prior to administering the composition to the subject. In some embodiments, absence of CNS lymphoma or CNS metastases is based on a lumbar puncture performed within 1 month prior to administering the composition to the subject.

[0036] In some embodiments, a subject does not have a history of Richter’s transformation of chronic leukemic lymphoma, small lymphocytic lymphoma, or lymphoplasmacytic lymphoma. In some embodiments, a subject does not have a fungal, bacterial, viral, or other infection that is uncontrolled or requiring intravenous (IV) antimicrobials for management. In some embodiments, a subject does not have acute symptoms of COVID- 19 infection. In some embodiments, a subject does not have a known history of infection with HIV, hepatitis B virus, and / or hepatitis C virus. In some embodiments, a subject does not have an active autoimmune disease and / or any other disease requiring immunosuppressive therapy or corticosteroid therapy. In some embodiments, a subject has not received greater than 10 mg / day of prednisone or equivalent dose of another corticosteroid. In some embodiments, a subject does not have a primary immunodeficiency or a history of a concomitant genetic syndrome associated with bone marrow failure. In some embodiments, a primary Page 7 of 173 12403779v1Attorney Docket No. 2017428-0684 immunodeficiency or the concomitant genetic syndrome associated with bone marrow failure comprises Fanconi anemia, Kostmann syndrome, or Shwachman-Diamond syndrome.

[0037] In some embodiments, a subject has not been treated with a systemic cancer therapy within 14 days or five half-lives of administration of the composition. In some embodiments, a subject has not been treated with a systemic cancer therapy comprising a biologic therapy within 28 days of administration of the composition. In some embodiments, a subject has not been treated with any investigational product within 28 days of administration of the composition. In some embodiments, a subject has not been treated with radiotherapy within 14 days of administration of the composition. In some embodiments, a subject has not been treated with major surgery within 28 days of administration of the composition. In some embodiments, a subject has not been treated with an autologous hematopoietic stem cell transplant (HSCT) within six weeks of administration of the composition. In some embodiments, a subject has not been treated with an allogeneic HSCT at any point prior to administration of the composition.

[0038] In some embodiments, a subject has not had a cardiac condition within one year of administration of the composition. In some embodiments, a cardiac condition comprises myocardial infarction, cardiac angioplasty or stenting, cardiac arrhythmia requiring medication, unstable angina, New York Heart Association Class II or greater congestive heart failure, cardiac atrial or ventricular lymphoma involvement, or a clinically significant cardiac disease. In some embodiments, a subject does not have a history of thromboembolic disease. In some embodiments, a subject does not have a history of non-line associated, clinically significant deep-vein thrombosis, proximal deep vein thrombosis, or pulmonary embolism requiring therapeutic anticoagulation within 6 months of administration of the composition.

[0039] In some embodiments, a subject does not have a history or presence of a CNS disorder within 12 months of administration of the composition. In some embodiments, a CNS disorder comprises a seizure disorder, a cerebrovascular ischemia / hemorrhage, a dementia, a cerebellar disease, or any autoimmune disease with CNS involvement.

[0040] In some embodiments, a subject does not have clinically significant ascites. In some embodiments, clinically significant ascites comprises physically positive ascites or ascites requiring puncture intervention or medication intervention for control.

[0041] In some embodiments, a subject does not have a history of organ transplantation and / or is not awaiting organ transplantation.

[0042] In some embodiments, a subject does not have an indwelling line or drain. In some embodiments, an indwelling line or drain comprises a percutaneous nephrostomy tube, an indwelling Foley catheter, a biliary drain, a G / J-tube, a pleural catheter, a peritoneal catheter, or a pericardial catheter.

[0043] In some embodiments, a subject has not received a live vaccine within 6 weeks of administration of the composition.

[0044] In some embodiments, a subject does not have an unresolved toxicity greater than Grade 1 from a previous anticancer therapy.

[0045] In some embodiments, a subject does not have a history of additional malignancy other than nonmelanoma skin cancer or carcinoma in situ. In some embodiments, a subject does not have a history of Page 8 of 173 12403779v1Attorney Docket No. 2017428-0684 additional malignancy, or has been disease-free from the additional malignancy for at least two years prior to administration of the composition.

[0046] In some embodiments, a subject does not have a severe immediate hypersensitivity to any of the components of the composition.

[0047] In some embodiments, a subject does not have an admission or evidence of illicit drug use, drug abuse, or alcohol abuse.

[0048] In some embodiments, a subject is 18-85 years of age. In some embodiments, a subject is 18-80 years of age. In some embodiments, a subject is 18-75 years of age.

[0049] In some embodiments, methods of the present disclosure further comprise: administering to a subject a lymphodepleting chemotherapy regimen, wherein the step of administering the lymphodepleting chemotherapy regimen occurs prior to the step of administering the composition. In some embodiments, a composition is administered 2-7 days after completion of the lymphodepleting chemotherapy regimen. In some embodiments, a composition is administered 3 days after completion of the lymphodepleting chemotherapy regimen. In some embodiments, a lymphodepleting chemotherapy is or comprises cyclophosphamide and fludarabine. In some embodiments, cyclophosphamide is administered at a dose of about 500 mg / m2. In some embodiments, cyclophosphamide is administered over approximately 60 minutes. In some embodiments, cyclophosphamide is administered daily for 3 days. In some embodiments, cyclophosphamide is administered to the subject 5 days, 4 days, and 3 days prior to the administration of the composition. In some embodiments, one liter of 0.9% sodium chloride is administered within 24 hours prior to administration of cyclophosphamide. In some embodiments, fludarabine is administered at a dose of about 30 mg / m2. In some embodiments, a subject has moderate impairment of renal function, and wherein fludarabine is administered at a dose of about 24 mg / m2with monitoring. In some embodiments, a subject has a creatinine clearance of about 50-70 mL / min / 1.73 m2. In some embodiments, fludarabine is administered over approximately 30 minutes. In some embodiments, fludarabine is administered daily for 3 days. In some embodiments, fludarabine is administered to the subject 5 days, 4 days, and 3 days prior to the administration of the composition. In some embodiments, one liter of 0.9% sodium chloride is administered at completion of administration of fludarabine. In some embodiments, cyclophosphamide and fludarabine are administered concurrently. In some embodiments, a lymphodepleting chemotherapy regimen is administered intravenously. In some embodiments, a lymphodepleting chemotherapy regimen further comprises administration of sodium 2-mercaptoethanesulfonate.

[0050] In some embodiments, methods of the present disclosure do not comprise administering to a subject one or more corticosteroids for at least 5 days prior to administration of the composition. In some embodiments, one or more corticosteroids comprise greater than or equal to 5 mg / day of prednisone or an equivalent dose of another corticosteroid. In some embodiments, a subject has computed tomography (CT) scans with contrast that are contraindicating for administration of systemic corticosteroids. In some embodiments, a subject undergoes MRI with contrast and noncontrast CT scans during the five days prior to administration of the composition. In some embodiments, methods of the present disclosure do not comprise administering to a subject one or more corticosteroids or other immunosuppressive drugs for at least three months after administration of the composition. In some embodiments, methods of the present disclosure comprise administering to a subject one or more corticosteroids or other immunosuppressive drugs after administration of the composition for management of composition-related toxicities. In some embodiments, Page 9 of 173 12403779v1Attorney Docket No. 2017428-0684 methods of the present disclosure do not comprise administering to the subject one or more corticosteroids, other immunosuppressive drugs, or other immunomodulatory medication at least one month prior to administration of the composition and at least two years after administration of the composition unless medically necessary. In some embodiments, an immunomodulatory medication comprises non-steroidal anti-inflammatory agents. In some embodiments, methods of the present disclosure do not comprise administering to a subject a lymphodepleting chemotherapy regimen prior to administration of the composition.

[0051] In some embodiments, methods of the present disclosure further comprise: administering to a subject a premedication therapy regimen, wherein a step of administering the premedication therapy regimen occurs prior to the step of administering a composition. In some embodiments, a premedication therapy is or comprises acetaminophen (paracetamol) and antihistamine. In some embodiments, a premedication therapy does not comprise corticosteroids or tocilizumab.

[0052] In some embodiments, methods of the present disclosure further comprise: administering to the subject a post-treatment therapy regimen, wherein a step of administering a post-treatment therapy regimen occurs after a step of administering a composition. In some embodiments, a post-treatment therapy is or comprises tocilizumab.

[0053] In some embodiments, methods of the present disclosure further comprise: collecting a sample from the subject after administration of the composition and determining: CAR T enumeration, CAR T cellular kinetics, CD47 expression, presence of replication competent lentivirus, T cell clonality, immunophenotype, and / or a cytokine profile.

[0054] In some embodiments, methods of the present disclosure further comprise modulating a subsequent administration of the composition, a post-treatment therapy, and / or one or more additional anti- cancer therapies based on the CAR T enumeration, CAR T cellular kinetics, CD47 expression, presence of replication competent lentivirus, T cell clonality, immunophenotype, and / or cytokine profile.

[0055] In some embodiments, methods of the present disclosure further comprise collecting a sample from a subject after administration of a composition and determining the presence of a: hypoimmunogenic T cell-mediated NK response, hypoimmunogenic T cell-mediated T response, hypoimmunogenic T cell-mediated humoral response, and / or humoral immunogenic response.

[0056] In some embodiments, methods of the present disclosure further comprise modulating a subsequent administration of the composition, a post-treatment therapy, and / or one or more additional anti- cancer therapies based on the presence of the hypoimmunogenic T cell-mediated NK response, hypoimmunogenic T cell-mediated T response, hypoimmunogenic T cell-mediated humoral response, and / or humoral immunogenic response.

[0057] In some embodiments, methods of the present disclosure further comprise: collecting a sample from the subject after administration of the composition and performing: single cell RNA sequencing, a minimal residual disease (MRD) assay a circulating tumor DNA (ctDNA) assay, and / or bulk RNA sequencing. In some embodiments, methods of the present disclosure further comprise modulating a subsequent administration of a composition, a post-treatment therapy, and / or one or more additional anti-cancer therapies based on the single cell RNA sequencing, an MRD assay, a ctDNA assay, or bulk RNA sequencing. Page 10 of 173 12403779v1Attorney Docket No. 2017428-0684

[0058] In some embodiments, methods of the present disclosure further comprise collecting a sample from a subject after administration of a composition and determining the presence of a cellular and / or humoral immune response directed towards one or more CD22-specific CAR peptides. In some embodiments, a cellular and / or humoral immune response is determined using ELISPOT or immunohistochemistry analysis. In some embodiments, methods of the present disclosure further comprise modulating a subsequent administration of a composition, a post-treatment therapy, and / or one or more additional anti-cancer therapies based on the presence of a cellular and / or humoral immune response.

[0059] In some embodiments, methods of the present disclosure further comprise: (i) obtaining first sequencing data for a first plurality of DNA molecules, (ii) determining, based on the first sequencing data, a first cancer profile, (iii) obtaining second sequencing data for a second plurality of DNA molecules, (iv) determining, based on the second sequencing data, a second cancer profile, and (v) detecting, based on a comparison of the first cancer profile and the second cancer profile a cancer status of the subject. In some embodiments, a cancer status is a minimal residual disease (MRD) status. In some embodiments, MRD status is based on identification of cancer specific markers in a first cancer profile and identification of the same cancer specific markers in a second cancer profile. In some embodiments, identification of cancer specific markers in a first cancer profile, a second cancer profile, or both, comprises comparing markers in the first cancer profile, the second cancer profile, or both to a reference cancer profile.

[0060] In some embodiments, a cancer status is a circulating tumor DNA (ctDNA) status. In some embodiments, ctDNA status is based on identification of cancer specific markers in a first cancer profile and identification of the same cancer specific markers in a second cancer profile. In some embodiments, identification of cancer specific markers in a first cancer profile, a second cancer profile, or both, comprises comparing the markers in the first cancer profile, the second cancer profile, or both to a reference cancer profile.

[0061] In some embodiments, methods of the present disclosure further comprise modulating a subsequent administration of a composition, a post-treatment therapy, and / or one or more additional anti-cancer therapies based in part on a change in the cancer status of the subject.

[0062] In some embodiments, methods of the present disclosure further comprise (i) obtaining first sequencing data for a first plurality of DNA molecules, (ii) determining, based on the first sequencing data, a first B-cell / T-cell profile for the patient, (iii) obtaining second sequencing data for a second plurality of DNA molecules, (iv) determining, based on the second sequencing data, a second B-cell / T-cell profile for the patient, (v) detecting, based on a comparison of the first B-cell / T-cell profile and the second B-cell / T-cell profile a change in the B-cell / T-cell status for a patient. In some embodiments, B-cell / T-cell status is the identity of B-cells and T- cells in the subject. In some embodiments, detection of the identity of the B-cells and T-cells in the subject is based on detection of rearranged IgH (VDJ) receptor gene sequences, rearranged IgH (DJ) receptor gene sequences, rearranged IgK receptor gene sequences, rearranged IgL receptor gene sequences, translocated BCL1 / IgH (J) receptor gene sequences, and / or translocated BCL2 / IgH(J) receptor gene sequences in the first B- cell / T-cell profile and detection of the same rearranged IgH (VDJ) receptor gene sequences, rearranged IgH (DJ) receptor gene sequences, rearranged IgK receptor gene sequences, rearranged IgL receptor gene sequences, translocated BCL1 / IgH (J) receptor gene sequences, and / or translocated BCL2 / IgH(J) receptor gene sequences in the second B-cell / T-cell profile. In some embodiments, detection of rearranged IgH (VDJ) receptor gene sequences, rearranged IgH (DJ) receptor gene sequences, rearranged IgK receptor gene sequences, rearranged IgL receptor gene sequences, translocated BCL1 / IgH (J) receptor gene sequences, and / or translocated Page 11 of 173 12403779v1Attorney Docket No. 2017428-0684 BCL2 / IgH(J) receptor gene sequences in the first B-cell / T-cell profile, the second B-cell / T-cell profile, or both, comprises comparing the detection of rearranged IgH (VDJ) receptor gene sequences, rearranged IgH (DJ) receptor gene sequences, rearranged IgK receptor gene sequences, rearranged IgL receptor gene sequences, translocated BCL1 / IgH (J) receptor gene sequences, and / or translocated BCL2 / IgH(J) receptor gene sequences in the first B-cell / T-cell profile, the second B-cell / T-cell profile, or both to a reference. In some embodiments, B- cell / T-cell status is the quantity of B-cells and T-cells in the subject. In some embodiments, detection of the quantity of the B-cells and T-cells in the subject is based on detection of rearranged IgH (VDJ) receptor gene sequences, rearranged IgH (DJ) receptor gene sequences, rearranged IgK receptor gene sequences, rearranged IgL receptor gene sequences, translocated BCL1 / IgH (J) receptor gene sequences, and / or translocated BCL2 / IgH(J) receptor gene sequences in the first B-cell / T-cell profile and detection of the same rearranged IgH (VDJ) receptor gene sequences, rearranged IgH (DJ) receptor gene sequences, rearranged IgK receptor gene sequences, rearranged IgL receptor gene sequences, translocated BCL1 / IgH (J) receptor gene sequences, and / or translocated BCL2 / IgH(J) receptor gene sequences in the second B-cell / T-cell profile. In some embodiments, detection of rearranged IgH (VDJ) receptor gene sequences, rearranged IgH (DJ) receptor gene sequences, rearranged IgK receptor gene sequences, rearranged IgL receptor gene sequences, translocated BCL1 / IgH (J) receptor gene sequences, and / or translocated BCL2 / IgH(J) receptor gene sequences in the first B-cell / T-cell profile, the second B-cell / T-cell profile, or both, comprises comparing the detection of rearranged IgH (VDJ) receptor gene sequences, rearranged IgH (DJ) receptor gene sequences, rearranged IgK receptor gene sequences, rearranged IgL receptor gene sequences, translocated BCL1 / IgH (J) receptor gene sequences, and / or translocated BCL2 / IgH(J) receptor gene sequences in the first B-cell / T-cell profile, the second B-cell / T-cell profile, or both to a reference.

[0063] In some embodiments, methods of the present disclosure further comprise modulating a subsequent administration of the composition, a post-treatment therapy, and / or one or more additional anti- cancer therapies based in part on a change in the subject’s status, the identity of B-cells and T-cells in the subject, the quantity of B-cells and T-cells in the subject, or a combination thereof.

[0064] In some embodiments, methods of the present disclosure further comprise (i) obtaining a first sample from the subject before administration of the composition; (ii) obtaining a first set of sequencing data for a first plurality of DNA molecules obtained from the first sample; (iii) determining a first cancer profile based on the first set of sequencing data; (iv) obtaining a second sample from the subject after administration of the composition; (v) obtaining a second set of sequencing data for a second plurality of DNA molecules obtained from the second sample; (vi) determining a second cancer profile based on the second set of sequencing data; and (vii) comparing the first cancer profile and the second cancer profile to determine a positive minimal residual disease (MRD) status or a negative MRD status in the subject.

[0065] In some embodiments, methods of the present disclosure further comprise (i) obtaining a first sample from the subject before administration of the composition; (ii) obtaining a first set of sequencing data for a first plurality of DNA molecules obtained from the first sample; (iii) determining a first B cell receptor (BCR) / TCR profile based on the first set of sequencing data; (iv) obtaining a second sample from the subject after administration of the composition; (v) obtaining a second set of sequencing data for a second plurality of DNA molecules obtained from the second sample; (vi) determining a second BCR / TCR profile based on the second set of sequencing data; and (vii) comparing the first BCR / TCR profile and the second BCR / TCR profile to determine a positive MRD status or a negative MRD status in the subject. Page 12 of 173 12403779v1Attorney Docket No. 2017428-0684

[0066] In some embodiments, methods of the present disclosure further comprise (i) obtaining a first sample from the subject before administration of the composition; (ii) obtaining a first set of sequencing data for a first plurality of DNA molecules obtained from the first sample; (iii) determining a first B cell / T cell profile based on the first set of sequencing data; (iv) obtaining a second sample from the subject after administration of the composition; (v) obtaining a second set of sequencing data for a second plurality of DNA molecules obtained from the second sample; (vi) determining a second B cell / T cell profile based on the second set of sequencing data; and (vii) comparing the first B cell / T cell profile and the second B Cell / T cell profile to determine a positive MRD status or negative MRD status in the subject.

[0067] In some embodiments, methods of the present disclosure further comprise (i) obtaining a first sample from the subject before administration of the composition; (ii) obtaining a first set of sequencing data for a first plurality of DNA molecules obtained from the first sample; (iii) determining a first B cell / T cell quantity based on the first set of sequencing data; (iv) obtaining a second sample from the subject after administration of the composition; (v) obtaining a second set of sequencing data for a second plurality of DNA molecules obtained from the second sample; (vi) determining a second B cell / T cell quantity based on the second set of sequencing data; and (vii) comparing the first B cell / T cell quantity and the second B Cell / T cell quantity to determine a positive MRD status or negative MRD status in the subject.

[0068] In some embodiments, if the subject is determined to have a positive MRD status, the subject is subsequently administered one or more additional anti-cancer therapies. In some embodiments, one or more additional anti-cancer therapies comprise a small molecule inhibitor, a chemotherapeutic agent, a cancer immunotherapy, an antibody, a cellular therapy, a nucleic acid, a surgery, a radiotherapy, an anti-angiogenic therapy, an anti-DNA repair therapy, an anti-inflammatory therapy, an anti-neoplastic agent, a growth inhibitory agent, a cytotoxic agent, or any combination thereof.

[0069] In some embodiments, methods of the present disclosure further comprise collecting a sample from a subject after administration of the composition and determining disease progression. In some embodiments, disease progression is determined by an increased diameter measurement of one or more malignant lesions as visualized via radiography. In some embodiments, a subject is administered one or more additional anti-cancer therapies in the presence of disease progression. In some embodiments, one or more additional anti-cancer therapies comprise a small molecule inhibitor, a chemotherapeutic agent, a cancer immunotherapy, an antibody, a cellular therapy, a nucleic acid, a surgery, a radiotherapy, an anti-angiogenic therapy, an anti-DNA repair therapy, an anti-inflammatory therapy, an anti-neoplastic agent, a growth inhibitory agent, a cytotoxic agent, or any combination thereof.

[0070] In some embodiments, engineered hypoimmunogenic T cells elicit reduced immune recognition when administered to the subject relative to the control or wild-type cells. In some embodiments, engineered hypoimmunogenic T cells elicit a reduced adaptive immune response when administered to the subject relative to the control or wild-type cells. In some embodiments, engineered hypoimmunogenic T cells elicit a reduced T cell response when administered to the subject relative to the control or wild-type cells. In some embodiments, engineered hypoimmunogenic T cells elicit a reduced innate immune response when administered to the subject relative to the control or wild-type cells. In some embodiments, engineered hypoimmunogenic T cells elicit a reduced natural killer (NK) cell response when administered to the subject relative to the control or wild-type cells. In some embodiments, engineered hypoimmunogenic T cells elicit a reduced macrophage response when administered to the subject relative to the control or wild-type cells. In some embodiments, engineered Page 13 of 173 12403779v1Attorney Docket No. 2017428-0684 hypoimmunogenic T cells elicit a reduced graft-versus-host reaction (GvHR) when administered to the subject relative to the control or wild-type cells. In some embodiments, engineered hypoimmunogenic T cells persist in the subject for a longer duration relative to the control or wild-type cells. In some embodiments, engineered hypoimmunogenic T cells have greater expansion in the subject relative to the control or wild-type cells.

[0071] In some embodiments, engineered hypoimmunogenic T cells have an improved: (i) objective response; (ii) duration of response; (iii) time to next treatment; (iv) progression-free survival; (v) event-free survival; (vi) overall survival; (vii) MRD negativity, or combination thereof, relative to an untreated subject and / or a subject treated with a standard of care therapy. In some embodiments, an event-free survival duration is at least about 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 months. In some embodiments, an event-free survival duration is about 10 months. In some embodiments, a progression-free survival duration is at least about 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 months. In some embodiments, a progression-free survival duration is about 15 months. In some embodiments, a complete response rate (CRR) is about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%. In some embodiments, a CRR is about 65%. In some embodiments, a CRR is about 50%. In some embodiments, a CRR is about 30%. In some embodiments, a durable CRR is at least about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%. In some embodiments, a durable CRR is about 30%. In some embodiments, a durable CRR is about 40%. In some embodiments, a durable CRR is about 50%. In some embodiments, a durable CRR is about 60%. In some embodiments, an overall response rate (ORR) is about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%. In some embodiments, an ORR is about 70%. In some embodiments, an ORR is about 80%. In some embodiments, a median duration of response (mDOR) is at least about 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 months. In some embodiments, an mDOR is at least about 12 months. In some embodiments, an MRD negativity is about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%. In some embodiments, an MRD negativity is about 70%. In some embodiments, a B cell malignancy is LBCL and the composition comprising a population of engineered hypoimmunogenic T cells is a second-line therapy. In some embodiments, a B cell malignancy is transplant-ineligible LBCL and the composition comprising a population of engineered hypoimmunogenic T cells is a second-line therapy. In some embodiments, a B cell malignancy is LBCL and the composition comprising a population of engineered hypoimmunogenic T cells is a third-line therapy. In some embodiments, a B cell malignancy is CLL and the composition comprising a population of engineered hypoimmunogenic T cells is a third-line therapy.

[0072] In some embodiments, methods of the present disclosure further comprise (i) obtaining a biological sample from a subject; and (ii) performing an analysis of a biological sample to determine, a CD19 expression status, a CD20 expression status, a CD22 expression status, or a combination thereof. In some embodiments, a biological sample was collected prior to the step of administering a composition. In some embodiments, a biological sample was collected after the step of administering the composition.

[0073] In some embodiments, a positive CD19 expression status, CD20 expression status, CD22 expression status, or combination thereof, is prognostic of an improved, (i) objective response; (ii) duration of response; (iii) time to next treatment; (iv) progression-free survival; (v) event-free survival; (vi) overall survival; or combination thereof, relative to the subject prior to administration of the composition.

[0074] In some embodiments, a subject is a human. Page 14 of 173 12403779v1Attorney Docket No. 2017428-0684

[0075] The present disclosure provides articles of manufacture. In some embodiments, articles of manufacture comprise a composition of a cell therapy, or one of a plurality of compositions of a cell therapy. In some embodiments, a composition of a cell therapy, or one of a plurality of compositions of a cell therapy comprises T cells expressing a CD22-directed chimeric antigen receptor (CAR). In some embodiments, articles of manufacture comprise instructions for administering a cell therapy. In some embodiments, instructions for administering a cell therapy specify administering a T cell composition according to methods of the present disclosure.

[0076] The present disclosure provides pharmaceutical compositions. In some embodiments, pharmaceutical compositions comprise a population of engineered hypoimmunogenic T cells. In some embodiments, a population of engineered hypoimmunogenic T cells comprises reduced expression of beta-2- microglobulin (B2M), Class II Major Histocompatability Complex Transactivator (CIITA), and T cell receptor alpha constant (TRAC) relative to a control or wild-type T cells. In some embodiments, a population of engineered hypoimmunogenic T cells comprises increased expression of CD47 encoded by a first exogenous polynucleotide relative to the control or wild-type T cells. In some embodiments, a population of engineered hypoimmunogenic T cells expression of a CD22-directed chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide. In some embodiments, pharmaceutical compositions of the present disclosure further comprise one or more excipients comprising: a cryopreservation medium; an isotonic electrolyte solution; or a combination thereof. In some embodiments, pharmaceutical compositions of the present disclosure further comprise dimethylsulfoxide (DMSO). In some embodiments, pharmaceutical compositions of the present disclosure further comprise human serum albumin (HSA). In some embodiments, an isotonic electrolyte solution comprises: sodium chloride; sodium gluconate; sodium acetate trihydrate; potassium chloride; magnesium chloride; or a combination thereof. In some embodiments, a pharmaceutical composition comprises a concentration of 7.5% DMSO (v / v) and 1.2% HSA (w / v). In some embodiments, a pharmaceutical composition comprises a concentration of 7.5% DMSO (v / v) and 0.3% HSA (w / v). BRIEF DESCRIPTION OF THE DRAWINGS

[0077] The drawings are for illustration purposes only, not for limitation.

[0078] FIG.1 shows a chart illustrating a study and subject treatment schema.

[0079] FIG.2 shows a chart illustrating exemplary grading and management of cytokine release syndrome.

[0080] FIG.3 shows a chart illustrating exemplary grading and management of immune effector cell- associated neurotoxicity syndrome.

[0081] FIG.4 shows graphs of NK cell and macrophage killing of CD22-directed CAR T cells (HLA-I / II KO, TCR KO, CD22-directed CAR / CD47 overexpression); WT CAR T Cells (HLA-I / II positive, CD22-directed CAR overexpression); Unedited T cells (HLA-I / II positive, unedited / untransduced) and TKO cells (HLA-I / II KO, TCR KO) in an xCELLigence NK and macrophage innate immune evasion assay.

[0082] FIG.5 is a bar graph showing IFNγ ELISPOT analysis of splenocytes collected from CD34+ humanized NSG SGM3 mice injected with CD22-directed CAR T cells WT CD22-directed T cells, or Unedited T cells, co-cultured with Mitomycin C-treated donor T cells. Page 15 of 173 12403779v1Attorney Docket No. 2017428-0684

[0083] FIG.6 are line graphs showing the in vitro cytotoxicity of CD22-directed CAR T cells of the present disclosure against CD19+ / CD22+, CD19- / CD22+, and CD19+ / CD22- NALM6 and RAJI tumor cells as measured using a luciferase assay.

[0084] FIG.7A are graphs showing the in vitro cytotoxicity of CD22-directed CAR T cells of the present disclosure against CD19+ / CD22+, CD19- / CD22+, and CD19+ / CD22- NALM6 tumor cells in an IncuCyte assay. FIG.7B are graphs showing T cell expansion in a 7-day IncuCyte assay where CD22-directed CAR T cells of the present disclosure are co-cultured with CD19+ / CD22+, CD19- / CD22+, or CD19+ / CD22- NALM6 tumor cells. FIG.7C are graphs showing the in vitro cytotoxicity of CD22-directed CAR T cells of the present disclosure against CD19+ / CD22+, CD19- / CD22+, and CD19+ / CD22- RAJI tumor cells in an IncuCyte assay. FIG.7D are graphs showing T cell expansion in a 7-day IncuCyte assay where CD22-directed CAR T cells of the present disclosure are co-cultured with CD19+ / CD22+, CD19- / CD22+, or CD19+ / CD22- RAJI tumor cells. FIG.7E are bar graphs of cytokine meso-scale discovery (MSD) analysis of 24 hr supernatant harvests of the cultures of FIGs.7A and 7B. FIG.7F are bar graphs of cytokine meso-scale discovery (MSD) analysis of 24 hr supernatant harvests of the cultures of FIGs.7C and 7D.

[0085] FIG.8A are line graphs showing anti-tumor activity of multi-donor CD22-directed CAR T cells of the present disclosure in a NALM6-Luc (CD19+ / CD22+) xenograft tumor model. FIG.8B is a bar graph showing the area under curve (AUC) of the graphs of FIG.8A.

[0086] FIG.9A is a line graph showing NALM6 CD19 escape tumor model flux analysis of CD22- directed T cells of the present disclosure. FIG.9B is a line graph showing RAJI CD19 escape tumor model flux analysis of CD22-directed T cells of the present disclosure.

[0087] FIG.10A is a diagram of the dosing schedule for a NALM6-Luc (CD19+ / CD22+) xenograft tumor model study. FIG.10B are graphs showing NALM6-Luc xenograft tumor growth in NSG mice administered with CD22-directed CAR T cells of the present disclosure by serial luminescent imaging using the Spectral Imaging System. FIG.10C are bar graphs of tumor cell and CAR T cell biodistribution in the bone marrow ad measured by flow cytometry. FIG. 10D are H&E-stained micrographs of skin and lung tissue sections in NSG mice administered with CD22-directed CAR T cells of the present disclosure. DETAILED DESCRIPTION I. INTRODUCTION

[0088] Described herein are engineered or modified immune evasive cells, including but not limited to human immune evasive cells. To overcome the problem of a subject's immune rejection of these primary and / or stem cell-derived transplants, the inventors have developed and describe herein hypoimmunogenic cells (e.g., hypoimmunogenic pluripotent cells, differentiated cells derived from such, and primary cells) that represent a viable source for any transplantable cell type. Such cells are protected from adaptive and / or innate immune rejection upon administration to a recipient subject. Advantageously, the cells disclosed herein are not rejected by the recipient subject's immune system, regardless of the subject's genetic make-up, as they are protected from adaptive and innate immune rejection upon administration to a recipient subject. In some embodiments, the engineered and / or hypoimmunogenic cells do not express major histocompatibility complex (MHC) class I and class II antigens and / or T-cell receptors. In certain embodiments, the engineered and / or hypoimmunogenic Page 16 of 173 12403779v1Attorney Docket No. 2017428-0684 cells do not express MHC I and II antigens and / or T-cell receptors and overexpress CD47 proteins. In certain embodiments, the engineered and / or hypoimmunogenic cells such as engineered and / or hypoimmunogenic T cells do not express MHC I and II antigens and / or T-cell receptors, overexpress CD47 proteins and express exogenous CARs.

[0089] In some embodiments, hypoimmunogenic cells outlined herein are not subject to an innate immune cell rejection. In some instances, hypoimmunogenic cells are not susceptible to NK cell-mediated lysis. In some instances, hypoimmunogenic cells are not susceptible to macrophage engulfment. In some embodiments, hypoimmunogenic cells are useful as a source of universally compatible cells or tissues (e.g., universal donor cells or tissues) that are transplanted into a recipient subject with little to no immunosuppressant agent needed. Such hypoimmunogenic cells retain cell-specific characteristics and features upon transplantation, including, e.g., pluripotency, as well as being capable of engraftment and functioning similarly to a corresponding native cell.

[0090] The technology disclosed herein utilizes expression of tolerogenic factors and modulation (e.g., reduction or elimination) of MHC I, MHC II, and / or TCR expression in human cells. In some embodiments, genome editing technologies utilizing rare-cutting endonucleases (e.g., CRISPR / Cas) are also used to reduce or eliminate expression of genes involved in an immune response (e.g., by deleting genomic DNA of genes involved in an immune response or by insertions of genomic DNA into such genes, such that gene expression is impacted) in the cells. In some embodiments, genome editing technologies or other gene modulation technologies are used to insert tolerance-inducing (tolerogenic) factors in human cells, rendering the cells and their progeny (include any differentiated cells prepared therefrom) able to evade immune recognition upon engrafting into a recipient subject. As such, the cells described herein exhibit modulated expression of one or more genes and factors that affect MHC I, MHC II, and / or TCR expression and evade the recipient subject’s immune system.

[0091] The genome editing techniques enable double-strand DNA breaks at desired locus sites. These controlled double-strand breaks promote homologous recombination at the specific locus sites. This process focuses on targeting specific sequences of nucleic acid molecules, such as chromosomes, with endonucleases that recognize and bind to the sequences and induce a double-stranded break in the nucleic acid molecule. The double-strand break is repaired either by an error-prone non-homologous end-joining (NHEJ) or by homologous recombination (HR).

[0092] The practice of the numerous embodiments will employ, unless indicated specifically to the contrary, conventional methods of chemistry, biochemistry, organic chemistry, molecular biology, microbiology, recombinant DNA techniques, genetics, immunology, and cell biology that are within the skill of the art, many of which are described below for the purpose of illustration. Such techniques are explained fully in the literature. See, e.g., Sambrook, et al., Molecular Cloning: A Laboratory Manual (3rd Edition, 2001); Sambrook, et al., Molecular Cloning: A Laboratory Manual (2nd Edition, 1989); Maniatis et al., Molecular Cloning: A Laboratory Manual (1982); Ausubel et al., Current Protocols in Molecular Biology (John Wiley and Sons, updated July 2008); Short Protocols in Molecular Biology: A Compendium of Methods from Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience; Glover, DNA Cloning: A Practical Approach, vol. I & II (IRL Press, Oxford, 1985); Anand, Techniques for the Analysis of Complex Genomes, (Academic Press, New York, 1992); Transcription and Translation (B. Hames & S. Higgins, Eds., 1984); Perbal, A Practical Guide to Molecular Cloning (1984); Harlow and Lane, Antibodies, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1998) Current Protocols in Immunology Q. E. Coligan, A. M. Kruisbeek, D. H. Margulies, E. M. Shevach and W. Strober, Page 17 of 173 12403779v1Attorney Docket No. 2017428-0684 eds., 1991); Annual Review of Immunology; as well as monographs in journals such as Advances in Immunology. II. DEFINITIONS

[0093] As described in the present disclosure, the following terms will be employed, and are defined as indicated below.

[0094] The term "autoimmune disease" refers to any disease or disorder in which the subject mounts an immune response against its own tissues and / or cells. Autoimmune disorders can affect almost every organ system in the subject (e.g., human), including, but not limited to, diseases of the nervous, gastrointestinal, and endocrine systems, as well as skin and other connective tissues, eyes, blood and blood vessels. Examples of autoimmune diseases include, but are not limited to Hashimoto's thyroiditis, Systemic lupus erythematosus, Sjogren's syndrome, Graves' disease, Scleroderma, Rheumatoid arthritis, Multiple sclerosis, Myasthenia gravis and Diabetes.

[0095] The term “best response rate” or “BOR” is defined as an incidence of subjects with a complete response, partial response, stable disease, and / or progressive disease.

[0096] The term "cancer" as used herein is defined as a hyperproliferation of cells whose unique trait (e.g., loss of normal controls) results in unregulated growth, lack of differentiation, local tissue invasion, and metastasis. With respect to the inventive methods, the cancer can be any cancer, including any of acute lymphocytic cancer, acute myeloid leukemia, alveolar rhabdomyosarcoma, bladder cancer, bone cancer, brain cancer, breast cancer, cancer of the anus, anal canal, or anorectum, cancer of the eye, cancer of the intrahepatic bile duct, cancer of the joints, cancer of the neck, gallbladder, or pleura, cancer of the nose, nasal cavity, or middle ear, cancer of the oral cavity, cancer of the vulva, chronic lymphocytic leukemia, chronic myeloid cancer, colon cancer, esophageal cancer, cervical cancer, fibrosarcoma, gastrointestinal carcinoid tumor, Hodgkin lymphoma, hypopharynx cancer, kidney cancer, larynx cancer, leukemia, liquid tumors, liver cancer, lung cancer, lymphoma, malignant mesothelioma, mastocytoma, melanoma, multiple myeloma, nasopharynx cancer, non- Hodgkin lymphoma, ovarian cancer, pancreatic cancer, peritoneum, omentum, and mesentery cancer, pharynx cancer, prostate cancer, rectal cancer, renal cancer, skin cancer, small intestine cancer, soft tissue cancer, solid tumors, stomach cancer, testicular cancer, thyroid cancer, ureter cancer, and / or urinary bladder cancer. As used herein, the term "tumor" refers to an abnormal growth of cells or tissues of the malignant type, unless otherwise specifically indicated and does not include a benign type tissue.

[0097] The term "chronic infectious disease" refers to a disease caused by an infectious agent wherein the infection has persisted. Such a disease may include hepatitis (A, B, or C), herpes virus (e.g., VZV, HSV-1, HSV-6, HSV-II, CMV, and EBV), and HIV / AIDS. Non-viral examples may include chronic fungal diseases such Aspergillosis, Candidiasis, Coccidioidomycosis, and diseases associated with Cryptococcus and Histoplasmosis. None limiting examples of chronic bacterial infectious agents may be Chlamydia pneumoniae, Listeria monocytogenes, and Mycobacterium tuberculosis. In some embodiments, the disorder is human immunodeficiency virus (HIV) infection. In some embodiments, the disorder is acquired immunodeficiency syndrome (AIDS). Page 18 of 173 12403779v1Attorney Docket No. 2017428-0684

[0098] As used herein, “clinically effective amount” refers to an amount sufficient to provide a clinical benefit in the treatment and / or management of a disease, disorder, or condition. In some embodiments, a clinically effective amount is an amount that has been shown to produce at least one improved clinical endpoint to the standard of care for the disease, disorder, or condition. In some embodiments, a clinically effective amount is an amount that has been demonstrated, for example in a clinical trial, to be sufficient to provide statistically significant and meaningful effectiveness for treating the disease, disorder, or condition. In some embodiments, the clinically effective amount is also a therapeutically effective amount. In other embodiments, the clinically effective amount is not a therapeutically effective amount.

[0099] In some embodiments, an alteration or modification (including, for example, genetic alterations or modifications) described herein results in reduced expression of a target or selected polynucleotide sequence. In some embodiments, an alteration or modification described herein results in reduced expression of a target or selected polypeptide sequence. In some embodiments, an alteration or modification described herein results in increased expression of a target or selected polynucleotide sequence. In some embodiments, an alteration or modification described herein results in increased expression of a target or selected polypeptide sequence.

[0100] The terms "decrease," "reduced," "reduction," and "decrease" are all used herein generally to mean a decrease by a statistically significant amount. However, for avoidance of doubt, decrease," "reduced," "reduction," "decrease" means a decrease by at least 10% as compared to a reference level, for example a decrease by at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% decrease (i.e. absent level as compared to a reference sample), or any decrease between 10-100% as compared to a reference level. In some embodiments, the cells are engineered to have reduced expression of one or more targets relative to an unaltered or unmodified wild-type cell.

[0101] The term “donor subject” refers to an animal, for example, a human from whom cells can be obtained. The “non-human animals” and “non-human mammals” as used interchangeably herein, includes mammals such as rats, mice, rabbits, sheep, cats, dogs, cows, pigs, and non-human primates. The term “donor subject” also encompasses any vertebrate including but not limited to mammals, reptiles, amphibians and fish. However, advantageously, the donor subject is a mammal such as a human, or other mammals such as a domesticated mammal, e.g., dog, cat, horse, and the like, or production mammal, e.g., cow, sheep, pig, and the like. A “donor subject” can also refer to more than one donor, for example one or more humans or non-human animals or non-human mammals.

[0102] The term “duration of response” or “DOR” is defined for subjects who experience a best objective response (complete response or partial response) and is the time from the first objective response to disease progression or death due to any cause, whichever occurs first. In some embodiments, if the progression or death is not observed, the DOR will be censored at the last evaluable assessment date on or prior to the censoring events defined as (i) ongoing without event, (ii) lost to follow-up, (iii) withdrew consent, (iv) new anticancer therapy (including stem cell transplant), and / or (v) adequate assessments no longer available.

[0103] The term "endogenous" refers to a referenced molecule or polypeptide that is naturally present in the cell. Similarly, the term when used in reference to expression of an encoding nucleic acid refers to expression of an encoding nucleic acid naturally contained within the cell and not exogenously introduced. Page 19 of 173 12403779v1Attorney Docket No. 2017428-0684 Similarly, the term when used in reference to a promoter sequence refers to a promoter sequence naturally contained within the cell and not exogenously introduced.

[0104] The term "engineered cell" as used herein refers to a cell that has been altered in at least some way by human intervention, including, for example, by genetic alterations or modifications such that the engineered cell differs from a wild-type cell.

[0105] The term “event-free survival” or “EFS” is the time from infusion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure to the earliest of date of death from any cause, disease progression or relapse, or new anti-cancer therapy (including stem cell transplant). In some embodiments, when a subject does not have any of the above events prior to data cutoff, EFS is censored at the last adequate response assessment date on or prior to the earliest censoring event (except for stem cell transplant). In some embodiments, the censoring reason could be (i) ongoing without event, (ii) lost to follow-up, (iii) withdrew consent, (iv) stem cell transplant (subjects who proceed to stem cell transplant after infusion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure will be censored at the time of stem cell transplant), and / or (v) event after at least 2 missing scheduled disease assessments.

[0106] As used herein, the term "exogenous" in the context of a polynucleotide or polypeptide being expressed is intended to mean that the referenced molecule or the referenced polypeptide is introduced into the cell of interest. The polypeptide can be introduced, for example, by introduction of an encoding nucleic acid into the genetic material of the cells such as by integration into a chromosome or as non-chromosomal genetic material such as a plasmid or expression vector. Therefore, the term as it is used in reference to expression of an encoding nucleic acid refers to introduction of the encoding nucleic acid in an expressible form into the cell. An exogenous polynucleotide can be inserted into at least one allele of the cell using viral transduction, for example, with a vector. In some embodiments, the vector is a pseudotyped, self-inactivating lentiviral vector that carries exogenous polynucleotide. In some embodiments, the vector is a self-inactivating lentiviral vector pseudotyped with a vesicular stomatitis VSV-G envelope, and which carries the exogenous polynucleotide. In some embodiments, the exogenous polynucleotide is inserted into at least one allele of the cell using viral transduction. In some embodiments, exogenous polynucleotide is inserted into at least one allele of the cell using a lentivirus based viral vector. In some embodiments, the exogenous polynucleotide is inserted into a safe harbor or target locus of at least one allele of the cell.

[0107] An "exogenous" molecule is a molecule, construct, factor and the like that is not normally present in a cell, but can be introduced into a cell by one or more genetic, biochemical or other methods. "Normal presence in the cell" is determined with respect to the particular developmental stage and environmental conditions of the cell. Thus, for example, a molecule that is present only during embryonic development of neurons is an exogenous molecule with respect to an adult neuron cell. An exogenous molecule can comprise, for example, a functioning version of a malfunctioning endogenous molecule or a malfunctioning version of a normally-functioning endogenous molecule.

[0108] An exogenous molecule or factor can be, among other things, a small molecule, such as is generated by a combinatorial chemistry process, or a macromolecule such as a protein, nucleic acid, carbohydrate, lipid, glycoprotein, lipoprotein, polysaccharide, any modified derivative of the above molecules, or any complex comprising one or more of the above molecules. Nucleic acids include DNA and RNA, can be single- or double-stranded; can be linear, branched or circular; and can be of any length. Nucleic acids include those Page 20 of 173 12403779v1Attorney Docket No. 2017428-0684 capable of forming duplexes, as well as triplex-forming nucleic acids. See, for example, U.S. Pat. Nos. 5,176,996 and 5,422,251. Proteins include, but are not limited to, DNA-binding proteins, transcription factors, chromatin remodeling factors, methylated DNA binding proteins, polymerases, methylases, demethylases, acetylases, deacetylases, kinases, phosphatases, integrases, recombinases, ligases, topoisomerases, gyrases and helicases.

[0109] An exogenous molecule or construct can be the same type of molecule as an endogenous molecule, e.g., an exogenous protein or nucleic acid. In such instances, the exogenous molecule is introduced into the cell at greater concentrations than that of the endogenous molecule in the cell. In some instances, an exogenous nucleic acid can comprise an infecting viral genome, a plasmid or episome introduced into a cell, or a chromosome that is not normally present in the cell. Methods for the introduction of exogenous molecules into cells are known to those of skill in the art and include, but are not limited to, lipid-mediated transfer (i.e., liposomes, including neutral and cationic lipids), electroporation, direct injection, cell fusion, particle bombardment, calcium phosphate co-precipitation, DEAE-dextran-mediated transfer and viral vector-mediated transfer.

[0110] A "gene," for the purposes of the present disclosure, includes a DNA region encoding a gene product, as well as all DNA regions which regulate the production of the gene product, whether or not such regulatory sequences are adjacent to coding and / or transcribed sequences. Accordingly, a gene includes, but is not necessarily limited to, promoter sequences, terminators, translational regulatory sequences such as ribosome binding sites and internal ribosome entry sites, enhancers, silencers, insulators, boundary elements, replication origins, matrix attachment sites and / or locus control regions.

[0111] "Gene expression" refers to the conversion of the information, contained in a gene, into a gene product. A gene product can be the direct transcriptional product of a gene (e.g., mRNA, tRNA, rRNA, antisense RNA, ribozyme, structural RNA or any other type of RNA) or a protein produced by translation of an mRNA. Gene products also include RNAs which are modified, by processes such as capping, polyadenylation, methylation, and editing, and proteins modified by, for example, methylation, acetylation, phosphorylation, ubiquitination, ADP- ribosylation, myristoylation, and / or glycosylation.

[0112] The term “genetic modification” and its grammatical equivalents as used herein can refer to one or more alterations of a nucleic acid, e.g., the nucleic acid within an organism's genome. For example, genetic modification can refer to alterations, additions, and / or deletion of genes or portions of genes or other nucleic acid sequences. A genetically modified cell can also refer to a cell with an added, deleted and / or altered gene or portion of a gene. A genetically modified cell can also refer to a cell with an added nucleic acid sequence that is not a gene or gene portion. Genetic modifications include, for example, both transient knock-in or knock- down mechanisms, and mechanisms that result in permanent knock-in, knock-down, or knock-out of target genes or portions of genes or nucleic acid sequences Genetic modifications include, for example, both transient knock-in and mechanisms that result in permanent knock-in of nucleic acids sequences Genetic modifications also include, for example, reduced or increased transcription, reduced or increased mRNA stability, reduced or increased translation, and reduced or increased protein stability.

[0113] As used herein, the terms "grafting", "administering," "introducing", "implanting" and "transplanting" as well as grammatical variations thereof are used interchangeably in the context of the placement of cells (e.g., cells described herein) into a subject, by a method or route which results in localization or at least partial localization of the introduced cells at a desired site or systemic introduction (e.g., into Page 21 of 173 12403779v1Attorney Docket No. 2017428-0684 circulation). The cells can be implanted directly to the desired site, or alternatively be administered by any appropriate route which results in delivery to a desired location in the subject where at least a portion of the implanted cells or components of the cells remain viable. The period of viability of the cells after administration to a subject can be as short as a few hours, e. g. twenty-four hours, to a few days, to as long as several years. In some embodiments, the cells can also be administered (e.g., injected) a location other than the desired site, such as in the brain or subcutaneously, for example, in a capsule to maintain the implanted cells at the implant location and avoid migration of the implanted cells.

[0114] By "HLA" or "human leukocyte antigen" complex is a gene complex encoding the MHC proteins in humans. These cell-surface proteins that make up the HLA complex are responsible for the regulation of the immune response to antigens. In humans, there are two MHCs, class I and class II, "HLA-I" and "HLA-II". HLA-I includes three proteins, HLA-A, HLA-B and HLA-C, which present peptides from the inside of the cell, and antigens presented by the HLA-I complex attract killer T-cells (also known as CD8+ T-cells or cytotoxic T cells). The HLA-I proteins are associated with β-2 microglobulin (B2M). HLA-II includes five proteins, HLA-DP, HLA-DM, HLA-DOB, HLA-DQ and HLA-DR, which present antigens from outside the cell to T lymphocytes. This stimulates CD4+ cells (also known as T-helper cells). It should be understood that the use of either "MHC" or "HLA" is not meant to be limiting, as it depends on whether the genes are from humans (HLA) or murine (MHC). Thus, as it relates to mammalian cells, these terms may be used interchangeably herein.

[0115] As used herein to characterize a cell, the term "hypoimmunogenic" generally means that such cell is less prone to innate or adaptive immune rejection by a subject into which such cells are transplanted, e.g., the cell is less prone to allorejection by a subject into which such cells are transplanted. For example, relative to a cell of the same cell type that does not comprise the modifications, such a hypoimmunogenic cell may be about 2.5%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97.5%, 99% or more less prone to innate or adaptive immune rejection by a subject into which such cells are transplanted. In some embodiments, genome editing technologies are used to modulate the expression of MHC I and MHC II genes, and thus, contribute to generation of a hypoimmunogenic cell. In some embodiments, a hypoimmunogenic cell evades immune rejection in an MHC-mismatched allogeneic recipient. In some instance, differentiated cells produced from the hypoimmunogenic stem cells outlined herein evade immune rejection when administered (e.g., transplanted or grafted) to an MHC-mismatched allogeneic recipient. In some embodiments, a hypoimmunogenic cell is protected from T cell-mediated adaptive immune rejection and / or innate immune cell rejection. Detailed descriptions of hypoimmunogenic cells, methods of producing thereof, and methods of using thereof are found in WO2016183041 filed May 9, 2015; WO2018132783 filed January 14, 2018; WO2018175390 filed March 20, 2018; WO2020018615 filed July 17, 2019; WO2020018620 filed July 17, 2019; PCT / US2020 / 44635 filed July 31, 2020; US62 / 881,840 filed August 1, 2019; US62 / 891,180 filed August 23, 2019; US63 / 016,190, filed April 27, 2020; and US63 / 052,360 filed July 15, 2020, the disclosures including the examples, sequence listings and figures are incorporated herein by reference in their entirety.

[0116] Hypoimmunogenicity of a cell can be determined by evaluating the immunogenicity of the cell such as the cell’s ability to elicit adaptive and innate immune responses or to avoid eliciting such adaptive and innate immune responses. Such immune response can be measured using assays recognized by those skilled in the art. In some embodiments, an immune response assay measures the effect of a hypoimmunogenic cell on T cell proliferation, T cell activation, T cell killing, donor specific antibody generation, NK cell proliferation, NK cell activation, and macrophage activity. In some cases, hypoimmunogenic cells and derivatives thereof undergo Page 22 of 173 12403779v1Attorney Docket No. 2017428-0684 decreased killing by T cells and / or NK cells upon administration to a subject. In some instances, the cells and derivatives thereof show decreased macrophage engulfment compared to an unmodified or wild-type cell. In some embodiments, a hypoimmunogenic cell elicits a reduced or diminished immune response in a recipient subject compared to a corresponding unmodified wild-type cell. In some embodiments, a hypoimmunogenic cell is nonimmunogenic or fails to elicit an immune response in a recipient subject.

[0117] The term percent "identity," in the context of two or more nucleic acid or polypeptide sequences, refers to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., BLASTP and BLASTN or other algorithms available to persons of skill) or by visual inspection. Depending on the application, the percent "identity" can exist over a region of the sequence being compared, e.g., over a functional domain, or, alternatively, exist over the full length of the two sequences to be compared. For sequence comparison, typically one sequence acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters.

[0118] Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math.2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol.48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see generally Ausubel et al., infra).

[0119] One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, which is described in Altschul et al., J. Mol. Biol. 215:403-410 (1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information.

[0120] "Immune signaling factor" as used herein refers to, in some cases, a molecule, protein, peptide and the like that activates immune signaling pathways.

[0121] "Immunosuppressive factor" or "immune regulatory factor" or "tolerogenic factor" as used herein include hypoimmunity factors, complement inhibitors, and other factors that modulate or affect the ability of a cell to be recognized by the immune system of a host or recipient subject upon administration, transplantation, or engraftment. These may be in combination with additional genetic modifications.

[0122] The terms "increased", "increase" or "enhance" or "activate" are all used herein to generally mean an increase by a statically significant amount; for the avoidance of any doubt, the terms "increased", "increase" or "enhance" or "activate" means an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold Page 23 of 173 12403779v1Attorney Docket No. 2017428-0684 increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level. In some embodiments, the reference level, also referred to as the basal level, is 0.

[0123] In some embodiments, the alteration is an indel. As used herein, "indel" refers to a mutation resulting from an insertion, deletion, or a combination thereof. As will be appreciated by those skilled in the art, an indel in a coding region of a genomic sequence will result in a frameshift mutation, unless the length of the indel is a multiple of three. In some embodiments, the alteration is a point mutation. As used herein, "point mutation" refers to a substitution that replaces one of the nucleotides. A gene editing (e.g., CRISPR / Cas) system of the present disclosure can be used to induce an indel of any length or a point mutation in a target polynucleotide sequence.

[0124] As used herein, “knock down” refers to a reduction in expression of the target mRNA or the corresponding target protein. Knock down is commonly reported relative to levels present following administration or expression of a noncontrol molecule that does not mediate reduction in expression levels of RNA (e.g., a non-targeting control shRNA, siRNA, or miRNA). In some embodiments, knock down of a target gene is achieved by genetic modification, including use of gene editing systems (e.g., CRISPR / Cas).

[0125] Knock down is commonly assessed by measuring the mRNA levels using quantitative polymerase chain reaction (qPCR) amplification or by measuring protein levels by western blot or enzyme-linked immunosorbent assay (ELISA). Analyzing the protein level provides an assessment of both mRNA cleavage as well as translation inhibition. Further techniques for measuring knock down include RNA solution hybridization, nuclease protection, northern hybridization, gene expression monitoring with a microarray, antibody binding, radioimmunoassay, and fluorescence activated cell analysis. Those skilled in the art will readily appreciate how to use the gene editing systems (e.g., CRISPR / Cas) of the present disclosure to knock out a target polynucleotide sequence or a portion thereof based upon the details described herein.

[0126] By "knock in" or “knock-in” herein is meant a genetic modification resulting from the insertion of a DNA sequence into a chromosomal locus in a host cell. This causes initiation of or increased levels of expression of the knocked in gene, portion of gene, or nucleic acid sequence inserted product, e.g., an increase in RNA transcript levels and / or encoded protein levels. As will be appreciated by those in the art, this can be accomplished in several ways, including inserting or adding one or more additional copies of the gene or portion thereof to the host cell or altering a regulatory component of the endogenous gene increasing expression of the protein is made or inserting a specific nucleic acid sequence whose expression is desired. This may be accomplished by modifying a promoter, adding a different promoter, adding an enhancer, adding other regulatory elements, or modifying other gene expression sequences.

[0127] As used herein, "knock out" or “knock-out” includes deleting all or a portion of a target polynucleotide sequence in a way that interferes with the translation or function of the target polynucleotide sequence. For example, a knock out can be achieved by altering a target polynucleotide sequence by inducing an insertion or a deletion (“indel”) in the target polynucleotide sequence, including in a functional domain of the target polynucleotide sequence (e.g., a DNA binding domain). Those skilled in the art will readily appreciate how to use the gene editing systems (e.g., CRISPR / Cas) of the present disclosure to knock out a target polynucleotide sequence or a portion thereof based upon the details described herein.

[0128] In some embodiments, a genetic modification or alteration results in a knock out or knock down of the target polynucleotide sequence or a portion thereof. Knocking out a target polynucleotide sequence Page 24 of 173 12403779v1Attorney Docket No. 2017428-0684 or a portion thereof using a gene editing system (e.g., CRISPR / Cas) of the present disclosure can be useful for a variety of applications. For example, knocking out a target polynucleotide sequence in a cell can be performed in vitro for research purposes. For ex vivo purposes, knocking out a target polynucleotide sequence in a cell can be useful for treating or preventing a disorder associated with expression of the target polynucleotide sequence (e.g., by knocking out a mutant allele in a cell ex vivo and introducing those cells comprising the knocked out mutant allele into a subject) or for changing the genotype or phenotype of a cell.

[0129] "Modulation" of gene expression refers to a change in the expression level of a gene. Modulation of expression can include, but is not limited to, gene activation and gene repression. Modulation may also be complete, i.e., wherein gene expression is totally inactivated or is activated to wild-type levels or beyond; or it may be partial, wherein gene expression is partially reduced, or partially activated to some fraction of wild- type levels.

[0130] The methods provided herein can be used to alter a target polynucleotide sequence in a cell. The present disclosure contemplates altering target polynucleotide sequences in a cell for any purpose. In some embodiments, the target polynucleotide sequence in a cell is altered to produce a mutant cell. As used herein, a "mutant cell" refers to a cell with a resulting genotype that differs from its original genotype. In some instances, a "mutant cell" exhibits a mutant phenotype, for example when a normally functioning gene is altered using the gene editing systems (e.g., CRISPR / Cas) systems of the present disclosure. In other instances, a "mutant cell" exhibits a wild-type phenotype, for example when a gene editing system (e.g., CRISPR / Cas) system of the present disclosure is used to correct a mutant genotype. In some embodiments, the target polynucleotide sequence in a cell is altered to correct or repair a genetic mutation (e.g., to restore a normal phenotype to the cell). In some embodiments, the target polynucleotide sequence in a cell is altered to induce a genetic mutation (e.g., to disrupt the function of a gene or genomic element).

[0131] The term “native cell” as used herein refers to a cell that is not otherwise modified (e.g., engineered). In some embodiments, a native cell is a naturally occurring wild-type or a control cell.

[0132] The term “objective response rate” or “ORR” is defined as a proportion of subjects with a best overall response of complete response or partial response as determined by an investigator and / or central assessment. Subjects who do not meet the criteria for an objective response by an analysis cutoff date will be considered non-responders. In some embodiments, disease assessments obtained after infusion and up through an observation of progression or start of new anti-cancer therapy will be used. In some embodiments, response assessment in non-Hodgkin lymphoma subjects will be based on the Lugano classification criteria and in chronic lymphocytic leukemia subjects based on the International Workshop on Chronic Lymphocytic Leukemia criteria.

[0133] The term "operatively linked" or "operably linked" are used interchangeably with reference to a juxtaposition of two or more components (such as sequence elements), in which the components are arranged such that both components function normally and allow the possibility that at least one of the components can mediate a function that is exerted upon at least one of the other components. By way of illustration, a transcriptional regulatory sequence, such as a promoter, is operatively linked to a coding sequence if the transcriptional regulatory sequence controls the level of transcription of the coding sequence in response to the presence or absence of one or more transcriptional regulatory factors. A transcriptional regulatory sequence is generally operatively linked in cis with a coding sequence, but need not be directly adjacent to it. For example, Page 25 of 173 12403779v1Attorney Docket No. 2017428-0684 an enhancer is a transcriptional regulatory sequence that is operatively linked to a coding sequence, even though they are not contiguous.

[0134] The term “overall survival” or “OS” is defined as the time from infusion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure to death from any cause. In some embodiments, subjects who have not died by the analysis data cutoff date will have survival time censored at their last date known to be alive. In some embodiments, for subjects alive or dead after the data cutoff date, survival time will be censored at the data cutoff date.

[0135] The term “progression free survival” or “PFS” is defined as the time of infusion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure to disease progression or death from any cause, whichever occurs first. In some embodiments, if the progression or death is not observed, the PFS will be censored at the last evaluable assessment date on or prior to the censoring events defined as (i) ongoing without event, (ii) lost to follow up, (iii) withdrew consent, (iv) new anticancer therapy (including stem cell transplant), and / or (v) adequate assessments no longer available.

[0136] As used herein, "promoter," "promoter sequence," or "promoter region" refers to a DNA regulatory region / sequence capable of binding RNA polymerase and involved in initiating transcription of a downstream coding or non-coding sequence. In some examples, the promoter sequence includes the transcription initiation site and extends upstream to include the minimum number of bases or elements necessary to initiate transcription at levels detectable above background. In some embodiments, the promoter sequence includes a transcription initiation site, as well as protein binding domains responsible for the binding of RNA polymerase. Eukaryotic promoters will often, but not always, contain "TATA" boxes and "CAT" boxes.

[0137] In some embodiments, the engineered and hypoimmunogenic cells described are propagated from a primary T cell or a progeny thereof. As used herein, the term “propagated from a primary T cell or a progeny thereof” encompasses the initial primary T cell that is isolated from the donor subject and any subsequent progeny thereof. As used herein, the term “progeny” encompasses, e.g., a first-generation progeny, i.e., the progeny is directly derived from, obtained from, obtainable from or derivable from the initial primary T cell by, e.g., traditional propagation methods. The term “progeny” also encompasses further generations such as second, third, fourth, fifth, sixth, seventh, or more generations, i.e., generations of cells which are derived from, obtained from, obtainable from or derivable from the former generation by, e.g., traditional propagation methods. The term “progeny” also encompasses modified cells that result from the modification or alteration of the initial primary T cell or a progeny thereof.

[0138] The term “recipient patient” refers to an animal, for example, a human to whom treatment, including prophylactic treatment, with the cells as described herein, is provided. For treatment of those infections, conditions or disease states, which are specific for a specific animal such as a human patient, the term patient refers to that specific animal. The term “recipient patient” also encompasses any vertebrate including but not limited to mammals, reptiles, amphibians and fish. However, advantageously, the recipient patient is a mammal such as a human, or other mammals such as a domesticated mammal, e.g., dog, cat, horse, and the like, or production mammal, e.g., cow, sheep, pig, and the like.

[0139] As used herein, the terms "regulatory sequences," "regulatory elements," and "control elements" are interchangeable and refer to polynucleotide sequences that are upstream (5' non-coding sequences), within, or downstream (3' non-translated sequences) of a polynucleotide target to be expressed. Page 26 of 173 12403779v1Attorney Docket No. 2017428-0684 Regulatory sequences influence, for example but are not limited to, the timing of transcription, amount or level of transcription, RNA processing or stability, and / or translation of the related structural nucleotide sequence. Regulatory sequences may include activator binding sequences, enhancers, introns, polyadenylation recognition sequences, promoters, repressor binding sequences, stem-loop structures, translational initiation sequences, translation leader sequences, transcription termination sequences, translation termination sequences, primer binding sites, and the like. It is recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, nucleotide sequences of different lengths may have identical regulatory or promoter activity.

[0140] As used herein, a “target” can refer to a gene, a portion of a gene, a portion of the genome, or a protein that is subject to regulatable reduced expression by the methods described herein.

[0141] As used herein, “therapeutically effective amount” refers to an amount sufficient to provide a therapeutic benefit in the treatment and / or management of a disease, disorder, or condition. In some embodiments, a therapeutically effective amount is an amount sufficient to ameliorate, palliate, stabilize, reverse, slow, attenuate or delay the progression of a disease, disorder, or condition, or of a symptom or side effect of the disease, disorder, or condition. In some embodiments, the therapeutically effective amount is also a clinically effective amount. In other embodiments, the therapeutically effective amount is not a clinically effective amount.

[0142] The term “time to next treatment” or “TTNT” is defined as the time from infusion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure to the next anticancer treatment (including stem cell transplant) or death from any cause, whichever occurs first. In some embodiments, when subsequent anticancer treatment is not received (e.g., continuing on current treatment) and subjects are still alive by the analysis data cutoff date, subjects will be censored at their last available follow-up date before the cutoff date.

[0143] As used herein, the term "treating" and "treatment" includes administering to a subject a therapeutically or clinically effective amount of cells described herein so that the subject has a reduction in at least one symptom of the disease or an improvement in the disease, for example, beneficial or desired therapeutic or clinical results. For purposes of this technology, beneficial or desired therapeutic or clinical results include, but are not limited to, alleviation of one or more symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. Treating can refer to prolonging survival as compared to expected survival if not receiving treatment. Thus, one of skill in the art realizes that a treatment may improve the disease condition, but may not be a complete cure for the disease. In some embodiments, one or more symptoms of a condition, disease or disorder are alleviated by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% upon treatment of the condition, disease or disorder.

[0144] For purposes of this technology, beneficial or desired therapeutic or clinical results of disease treatment include, but are not limited to, alleviation of one or more symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. Page 27 of 173 12403779v1Attorney Docket No. 2017428-0684

[0145] A "vector" or "construct" is capable of transferring gene sequences to target cells. Typically, "vector construct," "expression vector," and "gene transfer vector," mean any nucleic acid construct capable of directing the expression of a gene of interest and which can transfer gene sequences to target cells. Thus, the term includes cloning, and expression vehicles, as well as integrating vectors. Methods for the introduction of vectors or constructs into cells are known to those of skill in the art and include, but are not limited to, lipid- mediated transfer (i.e., liposomes, including neutral and cationic lipids), electroporation, direct injection, cell fusion, particle bombardment, calcium phosphate co-precipitation, DEAE-dextran-mediated transfer and / or viral vector-mediated transfer.

[0146] In some embodiments, the cells are engineered to have reduced or increased expression of one or more targets relative to an unaltered or unmodified wild-type cell. In some embodiments, the cells are engineered to have constitutive reduced or increased expression of one or more targets relative to an unaltered or unmodified wild-type cell. In some embodiments, the cells are engineered to have regulatable reduced or increased expression of one or more targets relative to an unaltered or unmodified wild-type cell. In some embodiments, the cells comprise increased expression of CD47 relative to a wild-type cell or a control cell of the same cell type. By “wild-type” or “wt” or “control” in the context of a cell means any cell found in nature. Examples of wild type or control cells include primary cells and T cells found in nature. However, by way of example, in the context of an engineered cell, as used herein, “wild-type” or “control” can also mean an engineered cell that may contain nucleic acid changes resulting in reduced expression of MHC I and / or II and / or T-cell receptors, but did not undergo the gene editing procedures to result in overexpression of CD47 proteins. For example, as used herein, “wild-type” or “control” means an engineered cell that comprises reduced or knocked out expression of B2M, CIITA, and / or TRAC. Also as used herein, “wild-type” or “control” means an engineered cell that comprises reduced or knocked out expression of B2M, CIITA, TRAC, and / or TRBC. As used herein, “wild-type” or “control” also means an engineered cell that may contain nucleic acid changes resulting in overexpression of CD47 proteins, but did not undergo the gene editing procedures to result in reduced expression of MHC I and / or II and / or T-cell receptors. In the context of an iPSC or a progeny thereof, “wild-type” or “control” also means an iPSC or progeny thereof that may contain nucleic acid changes resulting in pluripotency but did not undergo the gene editing procedures of the present disclosure to achieve reduced expression of MHC I and / or II and / or T-cell receptors, and / or overexpression of CD47 proteins. In the context of a primary T cell or a progeny thereof, “wild-type” or “control” also means a primary T cell or progeny thereof that may contain nucleic acid changes resulting in reduced expression of MHC I and / or II and / or T-cell receptors, but did not undergo the gene editing procedures to result in overexpression of CD47 proteins. For example, as used herein, “wild-type” or “control” means a primary T cell or progeny thereof that comprises reduced or knocked out expression of B2M, CIITA, and / or TRAC. Also as used herein, “wild-type” or “control” means a primary T cell or progeny thereof that comprises reduced or knocked out expression of B2M, CIITA, TRAC, and / or TRBC. Also in the context of a primary T cell or a progeny thereof, “wild-type” or “control” also means a primary T cell or progeny thereof that may contain nucleic acid changes resulting in overexpression of CD47 proteins, but did not undergo the gene editing procedures to result in reduced expression of MHC I and / or II and / or T-cell receptors. In some embodiments, the cells are engineered to have regulatable reduced or increased expression of one or more targets relative to a cell of the same cell type that does not comprise the modifications. In some embodiments, the wild-type cell or the control cell is a starting material. In some embodiments, the starting material is a primary cell collected from a donor. In some embodiments, the starting material is a primary blood cell collected from a donor, e.g., via a leukopak. For example, unmodified T cells Page 28 of 173 12403779v1Attorney Docket No. 2017428-0684 obtained from a donor is a starting material that are considered wild-type or control cells as contemplated herein. In another example, an iPSC cell line starting material is a starting material that is considered a wild-type or control cell as contemplated herein. In some embodiments, the starting material is otherwise modified or engineered to have altered expression of one or more genes to generate the engineered cell.

[0147] It is noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only,” and the like in connection with the recitation of claim elements, or use of a “negative” limitation. As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure. Any recited method may be carried out in the order of events recited or in any other order that is logically possible. Although any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present disclosure, representative illustrative methods and materials are now described.

[0148] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this technology belongs. Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the present disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the present disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the present disclosure. Certain ranges are presented herein with numerical values being preceded by the term “about.” The term “about” is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating unrecited number may be a number, which, in the context presented, provides the substantial equivalent of the specifically recited number. The term about is used herein to mean plus or minus ten percent (10%) of a value. For example, “about 100” refers to any number between 90 and 110.

[0149] All publications, patents, and patent applications cited in this specification are incorporated herein by reference to the same extent as if each individual publication, patent, or patent application were specifically and individually indicated to be incorporated by reference. Furthermore, each cited publication, patent, or patent application is incorporated herein by reference to disclose and describe the subject matter in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the technology described herein is not entitled to antedate such publication by virtue of prior technology. Further, the dates of publication provided might be different from the actual publication dates, which may need to be independently confirmed.

[0150] Before the technology is further described, it is to be understood that this technology is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims. It should also be Page 29 of 173 12403779v1Attorney Docket No. 2017428-0684 understood that the headers used herein are not limiting and are merely intended to orient the reader, but the subject matter generally applies to the technology disclosed herein. III. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0151] In NHL, patients progressing on a CD19-directed CAR T therapy have significant unmet medical need with no curative therapies. In DLBCL, while the durable clinical efficacy of autologous CAR T cell therapies has been established, limitations of the technology include long manufacturing and “vein to vein” times (i.e., time from leukapheresis to CAR T cell treatment) lasting 2–4 weeks. This may lead to treatment delays and disease progression in advanced subjects. Further, the reliance on the patient’s T cells, which may be dysfunctional or limited in numbers, contributes to treatment variability and failure, and may be associated with contamination with malignant tumor cells.

[0152] Off-the-shelf, allogeneic CAR T cell therapies may offer advantages over autologous strategies, including batch manufacturing from healthy donors, which allows immediate access to therapy and the option of retreatment. Allogeneic CAR T cells derived from healthy donors may overcome the challenges of T cell dysfunction in malignancy, providing a more consistently potent product with avoidance of malignant contamination. Further, a batch of allogeneic CAR T cells can be administered to multiple subjects, allowing a standardized treatment to be manufactured at reduced cost and reducing the barriers to patient access that currently exist.

[0153] A primary limitation of allogeneic CAR T cell approaches is limited CAR T cell expansion and persistence due in part to adaptive and / or innate immune recognition, leading to host-mediated CAR T cell destruction and poor patient responses. Additionally, the recipient’s cells may appear “foreign” to the T cell receptors (TCRs) of the allogeneic CAR T cells, which may induce their activation and result in acute graft- versus-host disease (GvHD). Current allogeneic CAR T cell therapies in development for NHL have utilized gene editing to delete TCR, which can effectively prevent GvHD. However, significant host-versus-graft immune responses exists for most allogeneic CAR T cell therapies despite gene editing of HLA class I and / or II genes, which may hinder adaptive immune recognition but augments innate immune recognition, thereby preventing the expansion and persistence of the allogeneic CAR T cell. Thus, there remains a significant need to develop allogeneic CAR T cell therapies with enhanced immune evasion and improved expansion, persistence, and response durability. A. Methods of Treatment with Hypoimmunogenic T Cells

[0154] As is described in further detail herein, provided herein are methods for treating a patient with a disease, disorder, or condition through administration of hypoimmunogenic cells, particularly hypoimmunogenic T cells. As will be appreciated, for all the multiple embodiments described herein related to the timing and / or combinations of therapies, administration of the cells is accomplished by a method or route which results in at least partial localization of the introduced cells at a desired site. Cells can be infused, implanted, or transplanted directly to the desired site, or alternatively be administered by any appropriate route which results in delivery to a desired location in the subject where at least a portion of the implanted cells or components of the cells remain viable. Page 30 of 173 12403779v1Attorney Docket No. 2017428-0684

[0155] Provided herein are methods for treating a patient with a disease, disorder, or condition of the present disclosure, which includes administration of a population of hypoimmunogenic cells (e.g., primary T cells) to a subject, e.g., a human patient. For instance, a population of hypoimmunogenic primary T cells such as, but limited to, CD3+ T cells, CD4+ T cells, CD8+ T cells, naïve T cells, regulatory T (Treg) cells, non-regulatory T cells, Th1 cells, Th2 cells, Th9 cells, Th17 cells, T-follicular helper (Tfh) cells, cytotoxic T lymphocytes (CTL), effector T (Teff) cells, central memory T (Tcm) cells, effector memory T (Tem) cells, effector memory T cells that express CD45RA (TEMRA cells), tissue-resident memory (Trm) cells, virtual memory T cells, innate memory T cells, memory stem cell (Tsc), γΔ T cells, and any other subtype of T cell is administered to a patient to treat a disease, disorder, or condition of the present disclosure.

[0156] In some embodiments, pharmaceutical compositions as described herein are co-administered with a therapeutic agent that that binds to and / or interacts with one or more receptors selected from the group consisting of CD94, KIR2DL4, PD-1, an inhibitory NK cell receptor, and an activating NK receptor. In some instances, a therapeutic agent binds to a receptor on the surface of an NK cell, including one or more subpopulations of NK cells. In some embodiments, a therapeutic agent is selected from the group consisting of an antibody and fragments and variants thereof, an antibody mimetic, a small molecule, a blocking peptide, and a receptor antagonist. In some embodiments, pharmaceutical compositions as described herein are administered as part of a combination therapy comprising a Bruton tyrosine kinase (BTK) inhibitor. In some embodiments, a BTK inhibitor is selected from a group consisting of: ibrutinib, acalabrutinib, zanubrutinib, and pirtorutinib. 1. Dosing and Formulations

[0157] Any therapeutically effective amount of cells described herein can be included in a pharmaceutical composition of the present disclosure, depending on the indication being treated. Non-limiting examples of the cells include primary T cells. In some embodiments, a pharmaceutical composition includes at least about 1x102, 5x 102, 1x103, 5x103, 1x104, 5x104, 1x105, 5x105, 1x106, 5x106, 1x107, 5x107, 1x108, 5x108, 1x109, 5x109, 1x1010, or 5x1010cells. In some embodiments, the pharmaceutical composition includes up to about 1x102, 5x102, 1x103, 5x103, 1x104, 5x104, 1x105, 5x105, 1x106, 5x106, 1x107, 5x107, 1x108, 5x108, 1x109, 5x109, 1x1010, or 5x1010cells. In some embodiments, the pharmaceutical composition includes up to about 6.0 x 108cells. In some embodiments, the pharmaceutical composition includes up to about 8.0 x 108cells. In some embodiments, the pharmaceutical composition includes at least 30x106cells. In some embodiments, the pharmaceutical composition includes approximately 30x106cells. In some embodiments, the pharmaceutical composition includes at least 60x106cells. In some embodiments, the pharmaceutical composition includes approximately 60x106cells. In some embodiments, the pharmaceutical composition includes at least 90x106cells. In some embodiments, the pharmaceutical composition includes approximately 90x106cells. In some embodiments, the pharmaceutical composition includes at least 120x106cells. In some embodiments, the pharmaceutical composition includes approximately 120x106cells. In some embodiments, the pharmaceutical composition includes at least 200x106cells. In some embodiments, the pharmaceutical composition includes approximately 200x106cells. In some embodiments, the pharmaceutical composition includes at least about 1x102to 5x102, 5x102to 1x103, 1x103to 5x103, 5x103to 1x104, 1x104to 5x104, 5x104to 1x105, 1x105to 5x105, 5x105to 1x106, 1x106to 5x106, 5x106to 1x107, 1x107to 5x107, 5x107to 1x108, 1x108to 5x108, 5x108to 1x109, 1x109to 5x109, 5x109to 1x1010, or 1x1010to 5x1010cells. In some embodiments, the pharmaceutical composition includes from about 1.0x106to about 2.5x108cells. In some embodiments, the pharmaceutical composition includes from about 2.0x106to about 2.0x108cells. In some embodiments, a pharmaceutical Page 31 of 173 12403779v1Attorney Docket No. 2017428-0684 composition of the present disclosure comprises from about 30x106cells to about 200x106cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises from about 20x106cells to about 400x106cells.

[0158] In some embodiments, a pharmaceutical composition of the present disclosure comprises about 40 x 106, 41 x 106, 42 x 106, 43 x 106, 44 x 106, 45 x 106, 46 x 106, 47 x 106, 48 x 106, 49 x 106, 50 x 106, x x x1x103, 5x103, 1x104, 5x104, 1x105, 5x105, 1x106, 5x106, 1x107, 5x107, 1x108, 5x108, 1x109, 5x109, 1x1010, or Page 32 of 173 12403779v1Attorney Docket No. 2017428-0684 5x1010CAR+ cells. In some embodiments, the pharmaceutical composition includes up to about 1x102, 5x102, 1x103, 5x103, 1x104, 5x104, 1x105, 5x105, 1x106, 5x106, 1x107, 5x107, 1x108, 5x108, 1x109, 5x109, 1x1010, or 5x1010CAR+ cells. In some embodiments, the pharmaceutical composition includes up to about 6.0x108CAR+ cells. In some embodiments, the pharmaceutical composition includes up to about 8.0x108CAR+ cells. In some embodiments, the pharmaceutical composition includes at least 30x106CAR+ cells. In some embodiments, the pharmaceutical composition includes approximately 30x106CAR+ cells. In some embodiments, the pharmaceutical composition includes at least 60x106CAR+ cells. In some embodiments, the pharmaceutical composition includes approximately 60x106CAR+ cells. In some embodiments, the pharmaceutical composition includes at least 120x106CAR+ cells. In some embodiments, the pharmaceutical composition includes approximately 120x106CAR+ cells. In some embodiments, the pharmaceutical composition includes at least 200x106CAR+ cells. In some embodiments, the pharmaceutical composition includes approximately 200x106CAR+ cells. In some embodiments, the pharmaceutical composition includes at least about 1 x 102to 5 x 102, 5 x 102to 1 x 103, 1 x 103to 5 x 103, 5 x 103to 1 x 104, 1 x 104to 5 x 104, 5 x 104to 1 x 105, 1 x 105to 5 x 105, 5 x 105to 1 x 106, 1 x 106to 5 x 106, 5 x 106to 1 x 107, 1 x 107to 5 x 107, 5 x 107to 1 x 108, 1 x 108to 5 x 108, 5 x 108to 1 x 109, 1 x 109to 5 x 109, 5 x 109to 1 x 1010, or 1 x 1010to 5 x 1010CAR+ cells. In some embodiments, the pharmaceutical composition includes from about 1.0 x 106to about 2.5 x 108CAR+ cells. In some embodiments, the pharmaceutical composition includes from about 2.0 x 106to about 2.0 x 108CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises from about 30x106CAR+ cells to about 200x106CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 5x106CAR+ cells to at least about 400x106CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 20x106CAR+ cells to about 400x106CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises at least about 30x106CAR+ cells, at least about 60x106CAR+ cells, at least about 90x106CAR+ cells, at least about 120x106CAR+ cells, at least about 150x106CAR+ cells, at least about 200x106CAR+ cells, at least about 250x106cells, at least about 300x106cells, at least about 350x106cells, or at least about 400x106cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises no more than about 200x106CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises no more than about 250x106CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises no more than about 300x106CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises no more than about 350x106CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises no more than about 400x106CAR+ cells.

[0160] In some embodiments, a pharmaceutical composition of the present disclosure comprises fully edited hypoimmunogenic CAR+ cells as described herein. In some embodiments, fully edited hypoimmunogenic CAR+ cells comprise one or more modifications that: (a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules, (b) increase expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not comprise the modification, and (c) express a CD19-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide. In Page 33 of 173 12403779v1Attorney Docket No. 2017428-0684 some embodiments, a pharmaceutical composition of the present disclosure comprises partially edited CAR+ cells as described herein. In some embodiments, partially edited CAR+ cells are missing one or more modifications that: (a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and / or (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules, and / or (b) increase expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not comprise the modification, relative to fully edited hypoimmunogenic CAR+ cells as described herein. In some embodiments, partially edited CAR+ cells (i) are missing one or more modifications that inactivate or disrupt one or more alleles of one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, and (ii) comprise one or more modifications that inactivate one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, relative to fully edited hypoimmunogenic CAR+ cells as described herein. In some embodiments, partially edited CAR+ cells (i) comprise one or more modifications that inactivate or disrupt one or more alleles of one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, and (ii) are missing one or more modifications that inactivate one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, relative to fully edited hypoimmunogenic CAR+ cells as described herein. In some embodiments, partially edited CAR+ cells (i) are missing one or more modifications that inactivate or disrupt one or more alleles of one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, and (ii) are missing one or more modifications that inactivate one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, relative to fully edited hypoimmunogenic CAR+ cells as described herein. In some embodiments, a pharmaceutical composition of the present disclosure comprises fully edited hypoimmunogenic CAR+ cells and partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises 100% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 20% to about 40% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 30% to about 50% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 40% to about 60% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 50% to about 70% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 60% to about 80% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 70% to about 90% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure Page 34 of 173 12403779v1Attorney Docket No. 2017428-0684 comprises about 80% to about 99% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 1% to about 10% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 5% to about 10% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 5% to about 15% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 5% to about 20% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 10% to about 30% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 20% to about 40% partially edited CAR+ cells.

[0161] In some embodiments, a pharmaceutical composition has a volume of at least 1.0, 1.25, 1.5, 1.75, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, or 500 mL. In exemplary embodiments, a pharmaceutical composition has a volume of up to about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, or 500 mL. In exemplary embodiments, a pharmaceutical composition has a volume of about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, or 500 mL. In some embodiments, a pharmaceutical composition has a volume of from about 1-50 mL, 50- 100 mL, 100-150 mL, 150-200 mL, 200-250 mL, 250-300 mL, 300-350 mL, 350-400 mL, 400-450 mL, or 450-500 mL. In some embodiments, a pharmaceutical composition has a volume of from about 1-50 mL, 50-100 mL, 100-150 mL, 150-200 mL, 200-250 mL, 250-300 mL, 300-350 mL, 350-400 mL, 400-450 mL, or 450-500 mL. In some embodiments, a pharmaceutical composition has a volume of from about 1-10 mL, 10-20 mL, 20-30 mL, 30-40 mL, 40-50 mL, 50-60 mL, 60-70 mL, 70-80 mL, 70-80 mL, 80-90 mL, or 90-100 mL. In some embodiments, a pharmaceutical composition has a volume that ranges from about 1.25mL to about 100mL. In some embodiments, a pharmaceutical composition has a volume that ranges from about 5 mL to about 80 mL. In some embodiments, a pharmaceutical composition has a volume that ranges from about 10 mL to about 70 mL. In some embodiments, a pharmaceutical composition has a volume that ranges from about 10 mL to about 50 mL. In some embodiments, a pharmaceutical composition has a volume of at least about 7.5 mL, at least about 15 mL, at least about 30 mL, or at least about 50 mL.

[0162] In some embodiments, a specific amount / dosage regimen will vary depending on the weight, gender, age and health of a subject; the formulation, the biochemical nature, bioactivity, bioavailability and the side effects of the cells and the number and identity of the cells in the complete therapeutic regimen.

[0163] In some embodiments, a therapeutically effective dose or a clinically effective dose of the pharmaceutical composition includes about 1.0x105to about 2.5x108cells at a volume of about 10 mL to 50 mL and the pharmaceutical composition is administered as a single therapeutically effective dose or clinically effective dose. In some cases, the therapeutically effective dose or clinically effective dose includes about 1.0x105to about 2.5x108primary T cells described herein at a volume of about 10 mL to 50 mL. In some embodiments, a therapeutically effective dose or clinically effective dose includes about 1.0x105to about 2.5x108primary T cells as disclosed herein at a volume of about 10 mL to 50 mL. Page 35 of 173 12403779v1Attorney Docket No. 2017428-0684

[0164] In some embodiments, a therapeutically effective dose or a clinically effective dose of the pharmaceutical composition comprises a concentration of engineered hypoimmunogenic T cells of at least 1.25x106cells / mL. In some embodiments, a therapeutically effective dose or a clinically effective dose of the pharmaceutical composition comprises a concentration of engineered hypoimmunogenic T cells of at least 4.0x106cells / mL. In some embodiments, a therapeutically effective dose or a clinically effective dose of the pharmaceutical composition comprises a concentration of engineered hypoimmunogenic T cells of at least 1.2x106CAR+ cells / mL. In some embodiments, a therapeutically effective dose or a clinically effective dose of the pharmaceutical composition comprises a concentration of engineered hypoimmunogenic T cells of at least 4.0x106CAR+ cells / mL.

[0165] In some embodiments, CD22 specific (CD22) CAR-T cells described herein are administered to a subject at a dose of about 40 x 106to about 400 x 106(e.g., 40 x 106, 41 x 106, 42 x 106, 43 x 106, 44 x 106, 45 x 106, 46 x 106, 47 x 106, 48 x 106, 49 x 106, 50 x 106, 51 x 106, 52 x 106, 53 x 106, 54 x 106, 55 x 106, 56 x 106, 57 x 106, 58 x 106, 59 x 106, 60 x 106, 61 x 106, 62 x 106, 63 x 106, 64 x 106, 65 x 106, 66 x 106, 67 x 106, 68 x 106, 69 x 106, 70 x 106, 71 x 106, 72 x 106, 73 x 106, 74 x 106, 75 x 106, 76 x 106, 77 x 106, 78 x 106, 79 x 106, 80 x 106, 81 x 106, 82 x 106, 83 x 106, 84 x 106, 85 x 106, 86 x 106, 87 x 106, 88 x 106, 89 x 106, 90 x 106, 91 x 106, 92 x 106, 93 x 106, 94 x 106, 95 x 106, 96 x 106, 97 x 106, 98 x 106, 99 x 106, 100 x 106, 101 x 106, 102 x 106, 103 x 106, 104 x 106, 105 x 106, 106 x 106, 107 x 106, 108 x 106, 109 x 106, 110 x 106, 111 x 106, 112 x 106, 113 x 106, 114 x 106, 115 x 106, 116 x 106, 117 x 106, 118 x 106, 119 x 106, 120 x 106, 121 x 106, 122 x 106, 123 x 106, 124 x 106, 125 x 106, 126 x 106, 127 x 106, 128 x 106, 129 x 106, 130 x 106, 131 x 106, 132 x 106, 133 x 106, 134 x 106, 135 x 106, 136 x 106, 137 x 106, 138 x 106, 139 x 106, 140 x 106, 141 x 106, 142 x 106, 143 x 106, 144 x 106, 145 x 106, 146 x 106, 147 x 106, 148 x 106, 149 x 106, 150 x 106, 151 x 106, 152 x 106, 153 x 106, 154 x 106, 155 x 106, 156 x 106, 157 x 106, 158 x 106, 159 x 106, 160 x 106, 161 x 106, 162 x 106, 163 x 106, 164 x 106, 165 x 106, 166 x 106, 167 x 106, 168 x 106, 169 x 106, 170 x 106, 171 x 106, 172 x 106, 173 x 106, 174 x 106, 175 x 106, 176 x 106, 177 x 106, 178 x 106, 179 x 106, 180 x 106, 181 x 106, 182 x 106, 183 x 106, 184 x 106, 185 x 106, 186 x 106, 187 x 106, 188 x 106, 189 x 106, 190 x 106, 191 x 106, 192 x 106, 193 x 106, 194 x 106, 195 x 106, 196 x 106, 197 x 106, 198 x 106, 199 x 106, 200 x 106, 201 x 106, 202 x 106, 203 x 106, 204 x 106, 205 x 106, 206 x 106, 207 x 106, 208 x 106, 209 x 106, 210 x 106, 211 x 106, 212 x 106, 213 x 106, 214 x 106, 215 x 106, 216 x 106, 217 x 106, 218 x 106, 219 x 106, 220 x 106, 221 x 106, 222 x 106, 223 x 106, 224 x 106, 225 x 106, 226 x 106, 227 x 106, 228 x 106, 229 x 106, 230 x 106, 231 x 106, 232 x 106, 233 x 106, 234 x 106, 235 x 106, 236 x 106, 237 x 106, 238 x 106, 239 x 106, 240 x 106, 241 x 106, 242 x 106, 243 x 106, 244 x 106, 245 x 106, 246 x 106, 247 x 106, 248 x 106, 249 x 106, 250 x 106, 251 x 106, 252 x 106, 253 x 106, 254 x 106, 255 x 106, 256 x 106, 257 x 106, 258 x 106, 259 x 106, 260 x 106, 261 x 106, 262 x 106, 263 x 106, 264 x 106, 265 x 106, 266 x 106, 267 x 106, 268 x 106, 269 x 106, 270 x 106, 271 x 106, 272 x 106, 273 x 106, 274 x 106, 275 x 106, 276 x 106, 277 x 106, 278 x 106, 279 x 106, 280 x 106, 281 x 106, 282 x 106, 283 x 106, 284 x 106, 285 x 106, 286 x 106, 287 x 106, 288 x 106, 289 x 106, 290 x 106, 291 x 106, 292 x 106, 293 x 106, 294 x 106, 295 x 106, 296 x 106, 297 x 106, 298 x 106, 299 x 106, 300 x 106, 301 x 106, 302 x 106, 303 x 106, 304 x 106, 305 x 106, 306 x 106, 307 x 106, 308 x 106, 309 x 106, 310 x 106, 311 x 106, 312 x 106, 313 x 106, 314 x 106, 315 x 106, 316 x 106, 317 x 106, 318 x 106, 319 x 106, 320 x 106, 321 x 106, 322 x 106, 323 x 106, 324 x 106, 325 x 106, 326 x 106, 327 x 106, 328 x 106, 329 x 106, 330 x 106, 331 x 106, 332 x 106, 333 x 106, 334 x 106, 335 x 106, 336 x 106, 337 x 106, 338 x 106, 339 x 106, 340 x 106, 341 x 106, 342 x 106, 343 x 106, 344 x 106, 345 x 106, 346 x 106, 347 x 106, 348 x 106, 349 x 106, 350 x 106, 351 x 106, 352 x 106, 353 x 106, 354 x 106, 355 x 106, 356 x 106, 357 x 106, 358 x 106, 359 x 106, 360 x 106, 361 x 106, 362 x 106, 363 x 106, 364 x 106, 365 x 106, 366 x 106, 367 x 106, 368 x 106, 369 x 106, 370 x 106, Page 36 of 173 12403779v1Attorney Docket No. 2017428-0684 371 x 106, 372 x 106, 373 x 106, 374 x 106, 375 x 106, 376 x 106, 377 x 106, 378 x 106, 379 x 106, 380 x 106, 381 x 106, 382 x 106, 383 x 106, 384 x 106, 385 x 106, 386 x 106, 387 x 106, 388 x 106, 389 x 106, 390 x 106, 391 x 106, 392 x 106, 393 x 106, 394 x 106, 395 x 106, 396 x 106, 397 x 106, 398 x 106, 399 x 106, or 400 x 106) viable CD22 specific CAR-T cells. In some embodiments, a dose is a therapeutically effective amount of viable CD22 specific CAR-T cells. In some embodiments, the dose is a clinically effective amount of viable CD22 specific CAR- T cells. In some embodiments, viable CD22 specific CAR-T cells include CD22 specific CAR expressing CD4+ T cells and CD22 specific CAR expressing CD8+ T cells at a ratio of about 1:1.

[0166] In some embodiments, a single dose of any of the CD22 specific (CD22) CAR-T cells described herein includes about 40 x 106to about 400 x 106(e.g., 40 x 106, 41 x 106, 42 x 106, 43 x 106, 44 x 106, 45 x x x x12403779v1Attorney Docket No. 2017428-0684 392 x 106, 393 x 106, 394 x 106, 395 x 106, 396 x 106, 397 x 106, 398 x 106, 399 x 106, or 400 x 106) viable CD22 specific CAR-T cells. In some embodiments, a dose is a therapeutically effective amount of viable CD22 specific CAR-T cells. In some embodiments, the dose is a clinically effective amount of viable CD22 specific CAR-T cells. In some embodiments, viable CD22 specific CAR-T cells include CD22 specific CAR expressing CD4+ T cells and CD22 specific CAR expressing CD8+ T cells at a ratio of about 1:1.

[0167] In some embodiments, the pharmaceutical composition is administered as a single therapeutically effective dose or clinically effective dose of from about 1.0x105to about 1.0x107cells (such as primary T cells) per kg body weight for subjects 50 kg or less. In some embodiments, the pharmaceutical composition is administered as a single therapeutically effective dose or clinically effective dose of from about 0.5x105to about 1.0x107, about 1.0x105to about 1.0x107, about 1.0x105to about 1.0x107, about 5.0x105to about 1x107, about 1.0x106to about 1x107, about 5.0x106to about 1.0x107, about 1.0x105to about 5.0x106, about 1.0x105to about 1.0x106, about 1.0x105to about 5.0x105, about 1.0x105to about 5.0x106, about 2.0x105to about 5.0x106, about 3.0x105to about 5.0x106, about 4.0x105to about 5.0x106, about 5.0x105to about 5.0x106, about 6.0x105to about 5.0x106, about 7.0x105to about 5.0x106, about 8.0x105to about 5.0x106, or about 9.0x105to about 5.0x106cells per kg body weight for subjects 50 kg or less. In some embodiments, the therapeutically effective dose or clinically effective dose is 0.5 x 105, 0.6 x 105, 0.7 x 105, 0.8 x 105, 0.9 x 105, 1.0 x 105, 1.1 x 105, 1.2 x 105, 1.3 x 105, 1.4 x 105, 1.5 x 105, 1.6 x 105, 1.7 x 105, 1.8 x 105, 1.9 x 105, 2.0 x 105, 2.1 x 105, 2.2 x 105, 2.3 x 105, 2.4 x 105, 2.5 x 105, 2.6 x 105, 2.7 x 105, 2.8 x 105, 2.9 x 105, 3.0 x 105, 3.1 x 105, 3.2 x 105, 3.3 x 105, 3.4 x 105, 3.5 x 105, 3.6 x 105, 3.7 x 105, 3.8 x 105, 3.9 x 105, 4.0 x 105, 4.1 x 105, 4.2 x 105, 4.3 x 105, 4.4 x 105, 4.5 x 105, 4.6 x 105, 4.7 x 105, 4.8 x 105, 4.9 x 105, 5.0 x 105, 0.5 x 106, 0.6 x 106, 0.7 x 106, 0.8 x 106, 0.9 x 106, 1.0 x 106, 1.1 x 106, 1.2 x 106, 1.3 x 106, 1.4 x 106, 1.5 x 106, 1.6 x 106, 1.7 x 106, 1.8 x 106, 1.9 x 106, 2.0 x 106, 2.1 x 106, 2.2 x 106, 2.3 x 106, 2.4 x 106, 2.5 x 106, 2.6 x 106, 2.7 x 106, 2.8 x 106, 2.9 x 106, 3.0 x 106, 3.1 x 106, 3.2 x 106, 3.3 x 106, 3.4 x 106, 3.5 x 106, 3.6 x 106, 3.7 x 106, 3.8 x 106, 3.9 x 106, 4.0 x 106, 4.1 x 106, 4.2 x 106, 4.3 x 106, 4.4 x 106, 4.5 x 106, 4.6 x 106, 4.7 x 106, 4.8 x 106, 4.9 x 106, 5.0 x 106, 5.1 x 106, 5.2 x 106, 5.3 x 106, 5.4 x 106, 5.5 x 106, 5.6 x 106, 5.7 x 106, 5.8 x 106, 5.9 x 106, 6.0 x 106, 6.1 x 106, 6.2 x 106, 6.3 x 106, 6.4 x 106, 6.5 x 106, 6.6 x 106, 6.7 x 106, 6.8 x 106, 6.9 x 106, 7.0 x 106, 7.1 x 106, 7.2 x 106, 7.3 x 106, 7.4 x 106, 7.5 x 106, 7.6 x 106, 7.7 x 106, 7.8 x 106, 7.9 x 106, 8.0 x 106, 8.1 x 106, 8.2 x 106, 8.3 x 106, 8.4 x 106, 8.5 x 106, 8.6 x 106, 8.7 x 106, 8.8 x 106, 8.9 x 106, 9.0 x 106, 9.1 x 106, 9.2 x 106, 9.3 x 106, 9.4 x 106, 9.5 x 106, 9.6 x 106, 9.7 x 106, 9.8 x 106, 9.9 x 106, 0.5 x 107, 0.6 x 107, 0.7 x 107, 0.8 x 107, 0.9 x 107, or 1.0 x 107cells per kg body weight for subjects 50 kg or less. In some embodiments, a therapeutically effective dose or clinically effective dose is from about 0.2 x 106to about 5.0 x 106cells per kg body weight for subjects 50 kg or less. In certain embodiments, a therapeutically effective dose or clinically effective dose is at a range that is lower than from about 0.2 x 106to about 5.0 x 106cells per kg body weight for subjects 50 kg or less. In some embodiments, a single therapeutically effective dose or clinically effective dose is at a volume of about 10 mL to 50 mL. In some embodiments, a therapeutically effective dose or clinically effective dose is administered intravenously. In some embodiments, a therapeutically effective dose or clinically effective dose is administered to the nervous system of a subject. In some embodiments, the nervous system is the central nervous system (CNS). In some embodiments, the nervous system is the peripheral nervous system (PNS). In some embodiments, a therapeutically effective dose or clinically effective dose is administered intrathecally.

[0168] In exemplary embodiments, cells are administered in a single therapeutically effective dose of from about 1.0x106to about 5.0x108cells (such as primary T cells) for subjects above 50 kg. In some Page 38 of 173 12403779v1Attorney Docket No. 2017428-0684 embodiments, a pharmaceutical composition is administered as a single therapeutically effective dose or clinically effective dose of from about 0.5x106to about 1.0x109, about 1.0x106to about 1.0x109, about 1.0x106to about 1.0x109, about 5.0x106to about 1.0x109, about 1.0x107to about 1.0x109, about 5.0x107to about 1.0x109, about 1.0x106to about 5.0x107, about 1.0x106to about 1.0x107, about 1.0x106to about 5.0x107, about 1.0x107to about 5.0x108, about 2.0x107to about 5.0x108, about 3.0x107to about 5.0x108, about 4.0x107to about 5.0x108, about 5.0x107to about 5.0x108, about 6.0x107to about 5.0x108, about 7.0x107to about 5.0x108, about 8.0x107to about 5.0x108, or about 9.0x107to about 5.0x108cells per kg body weight for subjects 50 kg or less. In some embodiments, a therapeutically effective dose or clinically effective dose is 1.0 x 106, 1.1 x 106, 1.2 x 106, 1.3 x 106, 1.4 x 106, 1.5 x 106, 1.6 x 106, 1.7 x 106, 1.8 x 106, 1.9 x 106, 2.0 x 106, 2.1 x 106, 2.2 x 106, 2.3 x 106, 2.4 x 106, 2.5 x 106, 2.6 x 106, 2.7 x 106, 2.8 x 106, 2.9 x 106, 3.0 x 106, 3.1 x 106, 3.2 x 106, 3.3 x 106, 3.4 x 106, 3.5 x 106, 3.6 x 106, 3.7 x 106, 3.8 x 106, 3.9 x 106, 4.0 x 106, 4.1 x 106, 4.2 x 106, 4.3 x 106, 4.4 x 106, 4.5 x 106, 4.6 x 106, 4.7 x 106, 4.8 x 106, 4.9 x 106, 5.0 x 106, 5.1 x 106, 5.2 x 106, 5.3 x 106, 5.4 x 106, 5.5 x 106, 5.6 x 106, 5.7 x 106, 5.8 x 106, 5.9 x 106, 6.0 x 106, 6.1 x 106, 6.2 x 106, 6.3 x 106, 6.4 x 106, 6.5 x 106, 6.6 x 106, 6.7 x 106, 6.8 x 106, 6.9 x 106, 7.0 x 106, 7.1 x 106, 7.2 x 106, 7.3 x 106, 7.4 x 106, 7.5 x 106, 7.6 x 106, 7.7 x 106, 7.8 x 106, 7.9 x 106, 8.0 x 106, 8.1 x 106, 8.2 x 106, 8.3 x 106, 8.4 x 106, 8.5 x 106, 8.6 x 106, 8.7 x 106, 8.8 x 106, 8.9 x 106, 9.0 x 106, 9.1 x 106, 9.2 x 106, 9.3 x 106, 9.4 x 106, 9.5 x 106, 9.6 x 106, 9.7 x 106, 9.8 x 106, 9.9 x 106, 1.0 x 107, 1.1 x 107, 1.2 x 107, 1.3 x 107, 1.4 x 107, 1.5 x 107, 1.6 x 107, 1.7 x 107, 1.8 x 107, 1.9 x 107, 2.0 x 107, 2.1 x 107, 2.2 x 107, 2.3 x 107, 2.4 x 107, 2.5 x 107, 2.6 x 107, 2.7 x 107, 2.8 x 107, 2.9 x 107, 3.0 x 107, 3.1 x 107, 3.2 x 107, 3.3 x 107, 3.4 x 107, 3.5 x 107, 3.6 x 107, 3.7 x 107, 3.8 x 107, 3.9 x 107, 4.0 x 107, 4.1 x 107, 4.2 x 107, 4.3 x 107, 4.4 x 107, 4.5 x 107, 4.6 x 107, 4.7 x 107, 4.8 x 107, 4.9 x 107, 5.0 x 107, 5.1 x 107, 5.2 x 107, 5.3 x 107, 5.4 x 107, 5.5 x 107, 5.6 x 107, 5.7 x 107, 5.8 x 107, 5.9 x 107, 6.0 x 107, 6.1 x 107, 6.2 x 107, 6.3 x 107, 6.4 x 107, 6.5 x 107, 6.6 x 107, 6.7 x 107, 6.8 x 107, 6.9 x 107, 7.0 x 107, 7.1 x 107, 7.2 x 107, 7.3 x 107, 7.4 x 107, 7.5 x 107, 7.6 x 107, 7.7 x 107, 7.8 x 107, 7.9 x 107, 8.0 x 107, 8.1 x 107, 8.2 x 107, 8.3 x 107, 8.4 x 107, 8.5 x 107, 8.6 x 107, 8.7 x 107, 8.8 x 107, 8.9 x 107, 9.0 x 107, 9.1 x 107, 9.2 x 107, 9.3 x 107, 9.4 x 107, 9.5 x 107, 9.6 x 107, 9.7 x 107, 9.8 x 107, 9.9 x 107, 1.0 x 108, 1.1 x 108, 1.2 x 108, 1.3 x 108, 1.4 x 108, 1.5 x 108, 1.6 x 108, 1.7 x 108, 1.8 x 108, 1.9 x 108, 2.0 x 108, 2.1 x 108, 2.2 x 108, 2.3 x 108, 2.4 x 108, 2.5 x 108, 2.6 x 108, 2.7 x 108, 2.8 x 108, 2.9 x 108, 3.0 x 108, 3.1 x 108, 3.2 x 108, 3.3 x 108, 3.4 x 108, 3.5 x 108, 3.6 x 108, 3.7 x 108, 3.8 x 108, 3.9 x 108, 4.0 x 108, 4.1 x 108, 4.2 x 108, 4.3 x 108, 4.4 x 108, 4.5 x 108, 4.6 x 108, 4.7 x 108, 4.8 x 108, 4.9 x 108, or 5.0 x 108cells per kg body weight for subjects 50 kg or less. In certain embodiments, cells are administered in a single therapeutically effective dose or clinically effective dose of about 1.0 x 107to about 2.5 x 108cells for subjects above 50 kg. In some embodiments, cells are administered in a single therapeutically effective dose or clinically effective dose of a range that is less than about 1.0 x 107to about 2.5 x 108cells for subjects above 50 kg. In some embodiments, cells are administered in a single therapeutically effective dose or clinically effective dose of a range that is higher than about 1.0 x 107to about 2.5 x 108cells for subjects above 50 kg. In some embodiments, a dose is administered intravenously. In some embodiments, a therapeutically effective dose or clinically effective dose is administered to the nervous system of a subject. In some embodiments, the nervous system is the central nervous system (CNS). In some embodiments, the nervous system is the peripheral nervous system (PNS). In some embodiments, a therapeutically effective dose or clinically effective dose is administered intrathecally. In some embodiments, a single therapeutically effective dose or clinically effective dose is at a volume of about 10 mL to 50 mL. In some embodiments, a therapeutically effective dose or clinically effective dose is administered intravenously. In some embodiments, a therapeutically effective dose or clinically effective dose is administered to the nervous system of a subject. In some embodiments, the nervous Page 39 of 173 12403779v1Attorney Docket No. 2017428-0684 system is the central nervous system (CNS). In some embodiments, the nervous system is the peripheral nervous system (PNS). In some embodiments, a therapeutically effective dose or clinically effective dose is administered intrathecally.

[0169] In exemplary embodiments, the therapeutically effective dose or clinically effective dose is administered intravenously at a rate of about 1 to 50 mL per minute, 1 to 40 mL per minute, 1 to 30 mL per minute, 1 to 20 mL per minute, 10 to 20 mL per minute, 10 to 30 mL per minute, 10 to 40 mL per minute, 10 to 50 mL per minute, 20 to 50 mL per minute, 30 to 50 mL per minute, 40 to 50 mL per minute. In numerous embodiments, a pharmaceutical composition is stored in one or more infusion bags for intravenous administration. In some embodiments, a dose is administered completely in no more than 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 70 minutes, 80 minutes, 90 minutes, 120 minutes, 150 minutes, 180 minutes, 240 minutes, or 300 minutes.

[0170] In some embodiments, a single therapeutically effective dose or clinically effective dose of the pharmaceutical composition is present in a single infusion bag. In other embodiments, a single therapeutically effective dose or clinically effective dose of a pharmaceutical composition is divided into 2, 3, 4 or 5 separate infusion bags.

[0171] In some embodiments, cells described herein are administered in a plurality of doses such as 2, 3, 4, 5, 6 or more doses, wherein the plurality of doses together constitute a therapeutically effective dose or clinically effective dose regimen. In some embodiments, each dose of a plurality of doses is administered to the subject ranging from 1 to 24 hours apart. In some instances, a subsequent dose is administered from about 1 hour to about 24 hours (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or about 24 hours) after an initial or preceding dose. In some embodiments, each dose of a plurality of doses is administered to the subject ranging from about 1 day to 28 days apart. In some instances, a subsequent dose is administered from about 1 day to about 28 days (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or about 28 days) after an initial or preceding dose. In certain embodiments, each dose of a plurality of doses is administered to the subject ranging from 1 week to about 6 weeks apart. In certain instances, a subsequent dose is administered from about 1 week to about 6 weeks (e.g., about 1, 2, 3, 4, 5, or 6 weeks) after an initial or preceding dose. In several embodiments, each dose of a plurality of doses is administered to the subject ranging from about 1 month to about 12 months apart. In several instances, a subsequent dose is administered from about 1 month to about 12 months (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months) after an initial or preceding dose.

[0172] In some embodiments, a subject is administered a first dosage regimen at a first time point, and then subsequently administered a second dosage regimen at a second time point. In some embodiments, the first dosage regimen is the same as the second dosage regimen. In other embodiments, the first dosage regimen is different than the second dosage regimen. In some instances, the number of cells in the first dosage regimen and the second dosage regimen are the same. In some instances, the number of cells in the first dosage regimen and the second dosage regimen are different. In some cases, the number of doses of the first dosage regimen and the second dosage regimen are the same. In some cases, the number of doses of the first dosage regimen and the second dosage regimen are different. Page 40 of 173 12403779v1Attorney Docket No. 2017428-0684

[0173] In some embodiments, CD22 specific (CD22) CAR-T cells described herein are administered to a subject at a dose of about 15 x 106to about 225 x 106(e.g., 15 x 106, 16 x 106, 17 x 106, 18 x 106, 19 x 106, 20 x 106, 21 x 106, 22 x 106, 23 x 106, 24 x 106, 25 x 106, 26 x 106, 27 x 106, 28 x 106, 29 x 106, 30 x 106, 31 x 106, 32 x 106, 33 x 106, 34 x 106, 35 x 106, 36 x 106, 37 x 106, 38 x 106, 39 x 106, 40 x 106, 41 x 106, 42 x 106, 43 x 106, 44 x 106, 45 x 106, 46 x 106, 47 x 106, 48 x 106, 49 x 106, 50 x 106, 60 x 106, 70 x 106, 80 x 106, 90 x 106, 100 x 106, 110 x 106, 111 x 106, 112 x 106, 113 x 106, 114 x 106, 115 x 106, 116 x 106, 117 x 106, 118 x 106, 119 x 106, 120 x 106, 121 x 106, 122 x 106, 123 x 106, 124 x 106, 125 x 106, 126 x 106, 127 x 106, 128 x 106, 129 x 106, 130 x 106, 131 x 106, 132 x 106, 133 x 106, 134 x 106, 135 x 106, 136 x 106, 137 x 106, 138 x 106, 139 x 106, 140 x 106, 141 x 106, 142 x 106, 143 x 106, 144 x 106, 145 x 106, 146 x 106, 147 x 106, 148 x 106, 149 x 106, 150 x 106, 151 x 106, 152 x 106, 153 x 106, 154 x 106, 155 x 106, 156 x 106, 157 x 106, 158 x 106, 159 x 106, 160 x 106, 161 x 106, 162 x 106, 163 x 106, 164 x 106, 165 x 106, 166 x 106, 167 x 106, 168 x 106, 169 x 106, 170 x 106, 171 x 106, 172 x 106, 173 x 106, 174 x 106, 175 x 106, 176 x 106, 177 x 106, 178 x 106, 179 x 106, 180 x 106, 181 x 106, 182 x 106, 183 x 106, 184 x 106, 185 x 106, 186 x 106, 187 x 106, 188 x 106, 189 x 106, 190 x 106, 191 x 106, 192 x 106, 193 x 106, 194 x 106, 195 x 106, 196 x 106, 197 x 106, 198 x 106, 199 x 106, 200 x 106, 201 x 106, 202 x 106, 203 x 106, 204 x 106, 205 x 106, 206 x 106, 207 x 106, 208 x 106, 209 x 106, 210 x 106, 211 x 106, 212 x 106, 213 x 106, 214 x 106, 215 x 106, 216 x 106, 217 x 106, 218 x 106, 219 x 106, 220 x 106, 221 x 106, 222 x 106, 223 x 106, 224 x 106, or 225 x 106) viable CD22 specific CAR-T cells. In some embodiments, a dose is a therapeutically effective amount of viable CD22 specific CAR-T cells. In some embodiments, the dose is a clinically effective amount of viable CD22 specific CAR-T cells. In some embodiments, viable CD22 specific CAR- T cells include CD22 specific CAR expressing CD4+ T cells and CD22 specific CAR expressing CD8+ T cells at a ratio of about 1:1.

[0174] In some embodiments, a subject is administered about 15 x 106to about 225 x 106(e.g., 15 x 16 x 17 x 18 x 19 x 20 x 21 x 22 x 23 x 24 x 25 x 26 x x T T TPage 41 of 173 12403779v1Attorney Docket No. 2017428-0684

[0175] In some embodiments, CD22 specific CAR-T cells as described herein are administered to a subject at a dose of about 0.1 x 106to 4 x 106(e.g., 0.1 x 106, 0.15 x 106, 0.2 x 106, 0.25 x 106, 0.3 x 106, 0.35 x 106, 0.4 x 106, 0.45 x 106, 0.5 x 106, 0.55 x 106, 0.6 x 106, 0.65 x 106, 0.7 x 106, 0.75 x 106, 0.8 x 106, 0.85 x 106, 0.9 x 106, 1.1 x 106, 1.15 x 106, 1.2 x 106, 1.25 x 106, 1.3 x 106, 1.35 x 106, 1.4 x 106, 1.45 x 106, 1.5 x 106, 1.55 x 106, 1.6 x 106, 1.65 x 106, 1.7 x 106, 1.75 x 106, 1.8 x 106, 1.85 x 106, 1.9 x 106, 1.95 x 106, 2 x 106, 2.1 x 106, 2.15 x 106, 2.2 x 106, 2.25 x 106, 2.3 x 106, 2.35 x 106, 2.4 x 106, 2.45 x 106, 2.5 x 106, 2.55 x 106, 2.6 x 106, 2.65 x 106, 2.7 x 106, 2.75 x 106, 2.8 x 106, 2.85 x 106, 2.9 x 106, 3 x 106, 3.1 x 106, 3.15 x 106, 3.2 x 106, 3.25 x 106, 3.3 x 106, 3.35 x 106, 3.4 x 106, 3.45 x 106, 3.5 x 106, 3.55 x 106, 3.6 x 106, 3.65 x 106, 3.7 x 106, 3.75 x 106, 3.8 x 106, 3.85 x 106, 3.9 x 106, 3.95 x 106, or 4 x 106) per kg of body weight. In some embodiments, a maximum dose administered is about 2 x 108viable CD22 specific CAR-T cells. In some embodiments, a maximum dose of about 2 x 108viable CD22 specific CAR-T cells is administered to a patient of about 100 kg of body weight and above. In some embodiments, the dose is a therapeutically effective amount of viable CD22 specific CAR-T cells. In other embodiments, the dose is a clinically effective amount of viable CD22 specific CAR-T cells.

[0176] In some embodiments, the CD22 specific CAR-T cells described herein are administered to a subject at a dose of up to about 2 x 108viable CD22 specific CAR-T cells. In some embodiments, a subject is administered from about 0.1 x 106to 4 x 106(e.g., 0.1 x 106, 0.15 x 106, 0.2 x 106, 0.25 x 106, 0.3 x 106, 0.35 x 106, 0.4 x 106, 0.45 x 106, 0.5 x 106, 0.55 x 106, 0.6 x 106, 0.65 x 106, 0.7 x 106, 0.75 x 106, 0.8 x 106, 0.85 x 106, 0.9 x 106, 1.1 x 106, 1.15 x 106, 1.2 x 106, 1.25 x 106, 1.3 x 106, 1.35 x 106, 1.4 x 106, 1.45 x 106, 1.5 x 106, 1.55 x 106, 1.6 x 106, 1.65 x 106, 1.7 x 106, 1.75 x 106, 1.8 x 106, 1.85 x 106, 1.9 x 106, 1.95 x 106, 2 x 106, 2.1 x 106, 2.15 x 106, 2.2 x 106, 2.25 x 106, 2.3 x 106, 2.35 x 106, 2.4 x 106, 2.45 x 106, 2.5 x 106, 2.55 x 106, 2.6 x 106, 2.65 x 106, 2.7 x 106, 2.75 x 106, 2.8 x 106, 2.85 x 106, 2.9 x 106, 3 x 106, 3.1 x 106, 3.15 x 106, 3.2 x 106, 3.25 x 106, 3.3 x 106, 3.35 x 106, 3.4 x 106, 3.45 x 106, 3.5 x 106, 3.55 x 106, 3.6 x 106, 3.65 x 106, 3.7 x 106, 3.75 x 106, 3.8 x 106, 3.85 x 106, 3.9 x 106, 3.95 x 106, or 4 x 106) viable CD22 specific CAR-T cells per kg of body weight for a subject with a body weight of about 50 kg or less. In some embodiments, a subject is administered from 0.1 x 106to 4 x 106(e.g., 0.1 x 106, 0.15 x 106, 0.2 x 106, 0.25 x 106, 0.3 x 106, 0.35 x 106, 0.4 x 106, 0.45 x 106, 0.5 x 106, 0.55 x 106, 0.6 x 106, 0.65 x 106, 0.7 x 106, 0.75 x 106, 0.8 x 106, 0.85 x 106, 0.9 x 106, 1.1 x 106, 1.15 x 106, 1.2 x 106, 1.25 x 106, 1.3 x 106, 1.35 x 106, 1.4 x 106, 1.45 x 106, 1.5 x 106, 1.55 x 106, 1.6 x 106, 1.65 x 106, 1.7 x 106, 1.75 x 106, 1.8 x 106, 1.85 x 106, 1.9 x 106, 1.95 x 106, 2 x 106, 2.1 x 106, 2.15 x 106, 2.2 x 106, 2.25 x 106, 2.3 x 106, 2.35 x 106, 2.4 x 106, 2.45 x 106, 2.5 x 106, 2.55 x 106, 2.6 x 106, 2.65 x 106, 2.7 x 106, 2.75 x 106, 2.8 x 106, 2.85 x 106, 2.9 x 106, 3 x 106, 3.1 x 106, 3.15 x 106, 3.2 x 106, 3.25 x 106, 3.3 x 106, 3.35 x 106, 3.4 x 106, 3.45 x 106, 3.5 x 106, 3.55 x 106, 3.6 x 106, 3.65 x 106, 3.7 x 106, 3.75 x 106, 3.8 x 106, 3.85 x 106, 3.9 x 106, 3.95 x 106, or 4 x 106) viable CD22 specific CAR-T cells for a subject with a body weight of greater than about 50 kg. In some embodiments, a subject is administered from about 0.2 x 108to about 2.0 x 108(e.g., about 0.2 x 108, 0.3 x 108, 0.4 x 108, 0.5 x 108, 0.6 x 108, 0.7 x 108, 0.8 x 108, 0.9 x 108, 1.0 x 108, 1.1 x 108, 1.2 x 108, 1.3 x 108, 1.4 x 108, 1.5 x 108, 1.6 x 108, 1.7 x 108, 1.8 x 108, 1.9 x 108, or 2.0 x 108) viable CD22 specific CAR-T cells. In some embodiments, the dose is a therapeutically effective amount of viable CD22 specific CAR-T cells. In other embodiments, the dose is a clinically effective amount of viable CD22 specific CAR-T cells.

[0177] In some embodiments, a single dose of any of the CD22 specific CAR-T cells described herein includes about 15 x 106to about 225 x 106(e.g., 15 x 106, 16 x 106, 17 x 106, 18 x 106, 19 x 106, 20 x 106, 21 x Page 42 of 173 12403779v1Attorney Docket No. 2017428-0684 106, 22 x 106, 23 x 106, 24 x 106, 25 x 106, 26 x 106, 27 x 106, 28 x 106, 29 x 106, 30 x 106, 31 x 106, 32 x 106, 33 x 106, 34 x 106, 35 x 106, 36 x 106, 37 x 106, 38 x 106, 39 x 106, 40 x 106, 41 x 106, 42 x 106, 43 x 106, 44 x 106, 45 x 106, 46 x 106, 47 x 106, 48 x 106, 49 x 106, 50 x 106, 60 x 106, 70 x 106, 80 x 106, 90 x 106, 100 x 106, 110 x 106, 111 x 106, 112 x 106, 113 x 106, 114 x 106, 115 x 106, 116 x 106, 117 x 106, 118 x 106, 119 x 106, 120 x 106, 121 x 106, 122 x 106, 123 x 106, 124 x 106, 125 x 106, 126 x 106, 127 x 106, 128 x 106, 129 x 106, 130 x 106, 131 x 106, 132 x 106, 133 x 106, 134 x 106, 135 x 106, 136 x 106, 137 x 106, 138 x 106, 139 x 106, 140 x 106, 141 x 106, 142 x 106, 143 x 106, 144 x 106, 145 x 106, 146 x 106, 147 x 106, 148 x 106, 149 x 106, 150 x 106, 151 x 106, 152 x 106, 153 x 106, 154 x 106, 155 x 106, 156 x 106, 157 x 106, 158 x 106, 159 x 106, 160 x 106, 161 x 106, 162 x 106, 163 x 106, 164 x 106, 165 x 106, 166 x 106, 167 x 106, 168 x 106, 169 x 106, 170 x 106, 171 x 106, 172 x 106, 173 x 106, 174 x 106, 175 x 106, 176 x 106, 177 x 106, 178 x 106, 179 x 106, 180 x 106, 181 x 106, 182 x 106, 183 x 106, 184 x 106, 185 x 106, 186 x 106, 187 x 106, 188 x 106, 189 x 106, 190 x 106, 191 x 106, 192 x 106, 193 x 106, 194 x 106, 195 x 106, 196 x 106, 197 x 106, 198 x 106, 199 x 106, 200 x 106, 201 x 106, 202 x 106, 203 x 106, 204 x 106, 205 x 106, 206 x 106, 207 x 106, 208 x 106, 209 x 106, 210 x 106, 211 x 106, 212 x 106, 213 x 106, 214 x 106, 215 x 106, 216 x 106, 217 x 106, 218 x 106, 219 x 106, 220 x 106, 221 x 106, 222 x 106, 223 x 106, 224 x 106, or 225 x 106) viable CD22 specific CAR-T cells. In some embodiments, the dose is a therapeutically effective amount of viable CD22 specific CAR-T cells. In other embodiments, the dose is a clinically effective amount of viable CD22 specific CAR-T cells. In some embodiments, the viable CD22 specific CAR-T cells include CD22 specific CAR expressing CD4+ T cells and CD22 specific CAR expressing CD8+ T cells at a ratio of about 1:1.

[0178] In some embodiments, a single dose of any of the CD22 specific CAR-T cells described herein includes about 2 x 108viable CD22 specific CAR-T cells. In some embodiments, a single infusion bag of any of the CD22 specific CAR-T cells described herein includes about 2 x 108viable CD22 specific CAR-T cells in a cell suspension of about 68 mL.

[0179] In some embodiments, a subject may be pre-medicated with a therapeutically effective amount of an antipyretic, an antihistamine, or an antipyretic and antihistamine prior to infusion of CD22-directed CAR-T cells of the present disclosure.

[0180] In some embodiments, an antipyretic includes acetaminophen, salicylamide, salicyl salicylate, methyl salicylate, magnesium salicylate, faislamine, ethenzamide, diflunisal, choline magnesium salicylate, benorylate / benorilatem and amoxiprin, acetylsalicylate, ceclofenac, acemetacin, alclofenac, bromfenac, diclofenac, etodolac, indomethacin, nabumetone, oxametacin, proglumetacin, sulindac, tolmetin, iminoprofen, benoxaprofen, carprofen, dexibuprofen, dexketoprofen, fenbufen, fenoprofen, flunoxaprofen, flurbiprofen, ibuprofen, ibuproxam, indoprofen, ketoprofen, ketorolac, loxoprofen, naproxen, oxaprozin, pirprofen, suprofen, tiaprofenic acid, mefenamic acid, flufenamic acid, meclofenamic acid, tolfenamic acid, droxicam, lornoxicam, meloxicam, piroxicam, tenoxicam, dipyrone, azapropazone, clofezone, kebuzone, metamizole, mofebutazone, oxyphenbutazone, phenazone, phenylbutazone, sulfinpyrazone, decoxib, rofecoxib, parecoxib, and etoricoxib. In some embodiments, the antipyretic is acetaminophen. In some embodiments, the therapeutically effective amount of acetaminophen is in the range of 300-1000 mg, 400-1000 mg, 500-1000 mg, 600-1000 mg, 700- 1000mg, 800-1000 mg, 900-1000 mg, 300-800 mg, 400-800 mg, 500-800 mg, 600-800 mg, 700-800 mg, 300- 600 mg, 400-600 mg, or 500-600 mg. In certain embodiments, the effective amount of acetaminophen is 300 mg, 400 mg.500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1000 mg. Page 43 of 173 12403779v1Attorney Docket No. 2017428-0684

[0181] In some embodiments, an antihistamine includes an H1 antihistamine, an H2 antihistamine, or an H1 and H2 antihistamine. In some embodiments, an antihistamine includes diphenhydramine, crivastine, azelastine, bilastine, brompheniramine, buclizine, bromodiphenhydramine, carbinoxamine, cetirizine, cyclizine, chlorpheniramine, chlorodiphenhydramine, clemastine, cromolyn, cyproheptadine, desloratadine, dexbrompheniramine, dexchlorpheniramine, dimenhydrinate, dimetindene, doxylamine, ebastine, embramine, fexofenadine, hydroxyzine, levocetirizine, loratadine, nedocromil, olopatadine, phenindamine, pheniramine, phenyltoloxamine, promethazine, pyrilamine, rupatadine, tripelennamine, or triprolidine. In some embodiments, the antihistamine is diphenhydramine. In certain embodiments, the therapeutically effective amount of diphenhydramine is 10 to100 mg, 20 to100 mg, 30 to100 mg, 40 to100 mg, 50 to100 mg, 60 to 100 mg, 70 to 100 mg, 80 to 100 mg, 90 to 100 mg, 10 to 90 mg, 20 to 90 mg, 30 to 90 mg, 40 to 90 mg, 50 to 90 mg, 60 to 90 mg, 70 to 90 mg, 80 to 90 mg, 10 to 80 mg, 20 to 80 mg, 30 to 80 mg, 40 to 80 mg, 50 to 80 mg, 60 to 80 mg, 70 to 80 mg, 10 to 70 mg, 20 to 70 mg, 30 to 70 mg, 40 to 70 mg, 50 to 70 mg, 60 to 70 mg, 10 to 60 mg, 20 to 60 mg, 30 to 60 mg, 40 to 60 mg, 50 to 60 mg, 10 to 50 mg, 20 to 50 mg, 30 to 50 mg, 40 to 50 mg, 20 to 40 mg, 30 to 40 mg, 20 to 30 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, or about 100 mg. 2. Pharmaceutical Compositions

[0182] For therapeutic application, cells prepared according to the disclosed methods can typically be supplied in the form of a pharmaceutical composition comprising an isotonic excipient and are prepared under conditions that are sufficiently sterile for human administration. For general principles in medicinal formulation of cell compositions, see "Cell Therapy: Stem Cell Transplantation, Gene Therapy, and Cellular Immunotherapy," by Morstyn & Sheridan eds, Cambridge University Press, 1996; and "Hematopoietic Stem Cell Therapy," E. D. Ball, J. Lister & P. Law, Churchill Livingstone, 2000. The cells can be packaged in a device or container suitable for distribution or clinical use.

[0183] In some embodiments, pharmaceutical compositions of the present disclosure comprise a population of engineered hypoimmunogenic T cells comprising: (a) reduced expression of Beta-2-Microglobulin (B2M), Class II Transactivator (CIITA), and T cell receptor alpha (TRAC) relative to a control T cell, (b) increased expression of CD47 encoded by a first exogenous polynucleotide relative to the control T cell, and (c) expression of a CD22-directed chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the population of engineered hypoimmunogenic T cells are allogeneic to the subject receiving the pharmaceutical composition. a. Pharmaceutically acceptable carriers

[0184] In some embodiments, a pharmaceutical composition provided herein further include a pharmaceutically acceptable carrier. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other Page 44 of 173 12403779v1Attorney Docket No. 2017428-0684 carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); salts such as sodium chloride; and / or non-ionic surfactants such as polysorbates (TWEEN™), poloxamers (PLURONICS™) or polyethylene glycol (PEG). In some embodiments, the pharmaceutical composition includes a pharmaceutically acceptable buffer (e.g., neutral buffer saline or phosphate buffered saline).

[0185] In some embodiments, a pharmaceutical composition includes one or more electrolyte base solutions selected from the group consisting of lactated CryoStor®, Ringer's solution, PlasmaLyte-A™, Iscove's Modified Dulbecco's Medium, Normosol-R™, Veen-D™, Polysal® and Hank's Balanced Salt Solution (containing no phenol red). These base solutions closely approximate the composition of extracellular mammalian physiological fluids.

[0186] In some embodiments, a pharmaceutical composition includes one or more cryoprotective agents selected from the group consisting of arabinogalactan, glycerol, polyvinylpyrrolidone (PVP), dextrose, dextran, trehalose, sucrose, raffinose, hydroxyethyl starch (HES), propylene glycol, human serum albumin (HSA), and dimethylsulfoxide (DMSO). In some embodiments, the pharmaceutically acceptable buffer is neutral buffer saline or phosphate buffered saline. In some embodiments, pharmaceutical compositions provided herein include one or more of CryoStor® CSB, Plasma-Lyte-A™, HSA, DMSO, and trehalose.

[0187] CryoStor® is an intracellular-like optimized solution containing osmotic / oncotic agents, free radical scavengers, and energy sources to minimize apoptosis, minimize ischemia / reperfusion injury and maximize the post-thaw recovery of the greatest numbers of viable, functional cells. CryoStor® is serum- and protein-free, and non-immunogenic. CryoStor® is cGMP-manufactured from raw materials of USP grade or higher. CryoStor® is a family of solutions pre-formulated with 0%, 2%, 5% or 10% DMSO. CryoStor® CSB is a DMSO-free version of CryoStor®. In some embodiments, a pharmaceutical composition includes a base solution of CryoStor® CSB at a concentration of about 0-100%, 5-95%, 10-90%, 15-85%, 20-80%, 30-80%, 40-80%, 50-80%, 60-80%, 70-80%, 25-75%, 30-70%, 35-65%, 40-60%, or 45-55% w / w. In some embodiments, a pharmaceutical composition includes a base solution of CryoStor® CSB at a concentration of about 0%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% w / w.

[0188] PlasmaLyte-A™ is a non-polymeric plasma expander and contains essential salts and nutrients similar to those found in culture medium but does not contain additional constituents found in tissue culture medium which are not approved for human infusion, e.g., phenol red, or are unavailable in U.S.P. grade. PlasmaLyte-A™ contains about 140 mEq / liter of sodium (Na), about 5 mEq / liter of potassium (K), about 3 mEq / liter of magnesium (Mg), about 98 mEq / liter of chloride (Cl), about 27 mEq / liter of acetate, and about 23 mEq / liter of gluconate. (PlasmaLyte-A™ is commercially available from Baxter, Hyland Division, Glendale Calif., product No.2B2543). In some embodiments, a pharmaceutical composition includes a base solution of PlasmaLyte-A™ at a concentration of about 0-100%, 5-95%, 10-90%, 15-85%, 15-80%, 15-75%, 15-70%, 15- 65%, 15-60%, 15-55%, 15-50%, 15-45%, 15-40%, 15-35%, 15-30%, 15-25%, 20-80%, 20-75%, 20-70%, 20- 65%, 20-60%, 20-55%, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 25-75%, 30-70%, 35-65%, 40-60%, or 45-55% w / w. In some embodiments, the pharmaceutical composition includes a base solution of PlasmaLyte-A™ at a concentration of about 0%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% w / w. Page 45 of 173 12403779v1Attorney Docket No. 2017428-0684

[0189] In some embodiments, a pharmaceutical composition includes a DMSO concentration (v / v) of at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 21%, at least about 22%, at least about 23%, at least about 24%, or at least about 25%. In some embodiments, a pharmaceutical composition includes a DMSO concentration (v / v) of about 7.5%.

[0190] In some embodiments, a pharmaceutical composition includes human serum albumin (HSA) at a concentration of about 0-10%, 0.3-9.3%, 0.3-8.3%, 0.3-7.3%, 0.3-6.3%, 0.3-5.3%, 0.3-4.3%, 0.3-3.3%, 0.3- 2.3%, 0.3-1.3%, 0.6-8.3%, 0.9-7.3%, 1.2-6.3%, 1.5-5.3%, 1.8-4.3%, or 2.1-3.3% w / v. In some embodiments, a pharmaceutical composition comprises human serum albumin (HSA) at a concentration (w / v) of at least about 0.1%, at least about 0.2%, at least about 0.3%, at least about 0.4%, at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 21%, at least about 22%, at least about 23%, at least about 24%, or at least about 25%. In some embodiments, a pharmaceutical composition includes HSA at a concentration of about 0%, 0.3%, 0.6%, 0.9%, 1.2%, 1.5%, 1.8%, 2.1%, 2.4%, 2.7%, 3.0%, 3.3%, 3.6%, 3.9%, 4.3%, 4.6%, 4.9%, 5.3%, 5.6%, 5.9%, 6.3%, 6.6%, 6.9%, 7.3%, 7.6%, 7.9%, 8.3%, 8.6%, 8.9%, 9.3%, 9.6%, 9.9%, or 10% w / v. In some embodiments, a pharmaceutical composition comprises HSA at a concentration (w / v) of about 0.3%.

[0191] In some embodiments, a pharmaceutical composition includes dimethyl sulfoxide (DMSO) at a concentration of about 0-10%, 0.5-9.5%, 1-9%, 1.5-8.5%, 2-8%, 3-8%, 4-8%, 5-8%, 6-8%, 7-8%, 2.5-7.5%, 3-7%, 3.5-6.5%, 4-6%, or 4.5-5.5% v / v. In some embodiments, a pharmaceutical composition includes HSA at a concentration of about 0%, 0.25%, 0.5%, 0.75%, 1.0%,1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%, 4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8.0%, 8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, or 10.0% v / v.

[0192] In some embodiments, a pharmaceutical composition includes dimethyl sulfoxide (DMSO) at a concentration of about 0-10%, 0.5-9.5%, 1-9%, 1.5-8.5%, 2-8%, 3-8%, 4-8%, 5-8%, 6-8%, 7-8%, 2.5-7.5%, 3-7%, 3.5-6.5%, 4-6%, or 4.5-5.5% v / v. In some embodiments, a pharmaceutical composition includes HSA at a concentration of about 0%, 0.25%, 0.5%, 0.75%, 1.0%,1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%, 4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8.0%, 8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, or 10.0% w / v.

[0193] In some embodiments, a pharmaceutical composition includes trehalose at a concentration of about 0-500 mM, 50-450 mM, 100-400 mM, 150-350 mM, or 200-300 mM. In some embodiments, a pharmaceutical composition includes trehalose at a concentration of about 0 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 125 mM, 150 mM, 175 mM, 200 mM, 225 mM, 250 mM, 275 mM, 300 mM, 325 mM, 350 mM, 375 mM, 400 mM, 425 mM, 450 mM, 475 mM, or 500 mM.

[0194] Exemplary pharmaceutical composition components are shown in TABLE 1. Page 46 of 173 12403779v1Attorney Docket No. 2017428-0684 TABLE 1: Exemplary pharmaceutical composition components Formulation Base Solution [DMSO] [HSA] [trehalose] A 7.5% 0.3% B 75% CroStor® CSB 3.75% 0.3% + 25% Pl L t(v / v) of 5% dextrose / 0.45% sodium chloride, 10% dextran 40 (LMD) / 5% dextrose, 20% (w / v) of 25% human serum albumin (HSA), and 7.5% (v / v) dimethyl sulfoxide (DMSO).

[0196] In some embodiments, a pharmaceutical composition comprises hypoimmunogenic cells described herein and a pharmaceutically acceptable carrier comprising 75% Cryostor CS10, 25% (v / v) Plasma- Lyte A, 7.5% (v / v) DMSO, and 1.2% (w / v) HSA. 3. Diseases, Disorders and Conditions to be Treated

[0197] B cell lymphomas encompass a group of lymphoma subtypes categorized within 2 broad categories: non-Hodgkin lymphoma (NHL) and Hodgkin lymphoma (HL), with NHLs being the majority of diagnosed cases. The therapeutic landscape varies for each distinct tumor subtype, but generally there are limited effective therapies in relapsed and / or refractory (r / r) settings across B cell lymphomas (National Comprehensive Cancer Network [NCCN] 2021). Despite recent advancement, NHL remains an unmet medical need. In 2021 American Cancer Society estimated that 82,000 cases of NHL would be diagnosed, with approximately 20,000 attributed deaths in the US in 2021 (American Cancer Society 2021). In 2020, there were 123,000 new cases of NHL reported, with 50,000 attributed deaths in Europe (World Health Organization 2020). NHL is a heterogeneous group of cancers originating in B lymphocytes, T lymphocytes, or natural killer (NK) cells. In the US, B cell lymphomas represent approximately 85% of all NHL cases (American Cancer Society 2019). Subtypes of NHL include large B cell lymphomas (LBCL), such as diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), and primary mediastinal B cell lymphoma (PMBCL), along with mantle lymphoma (MCL). Subtypes of indolent NHL (iNHL) include follicular lymphoma (FL) and marginal zone lymphoma (MZL).

[0198] DLBCL is the most aggressive NHL in adults, accounting for a third of all NHL cases, about 25,000 annually. In the US, the incidence of DLBCL is approximately 7 cases per 100,000 persons per year, with Caucasian-Americans having higher rates than other racial groups. There is a male predominance, and incidence increases with age with a median age of diagnosis of 64 years (Morton 2006, Shenoy 2011). Gene expression profiling has identified 2 distinct subtypes within DLBCL that are managed with similar standard of care— germinal center B cell (GCB) subtype and activated B cell (ABC) subtype. GCB subtype is associated with improved outcomes to therapy. MYC rearrangements are found in 5% to 8% of DLBCL patients and correlate with higher risk of disease progression (NCCN 2021).

[0199] HGBCL includes DLBCLs with MYC and B cell lymphoma 2 (BCL2) and / or B cell lymphoma 6 (BCL6) rearrangements (known as “double-hit” or “triple-hit lymphomas” based on the number of translocations). Generally, MYC rearrangements in DLBCLs are associated with a poor prognosis that is worsened in cases of concomitant BCL2 and / or BCL6 alterations. The majority of HGBCL are germinal center B Page 47 of 173 12403779v1Attorney Docket No. 2017428-0684 cell like lymphomas and present with worse outcomes than DLBCL, though treatment algorithms are shared. Exceptions to HGBCL include cases of follicular or lymphoblastic lymphoma or HGBCL, not otherwise specified (NOS), which contains blastoid-appearing LBCLs or cases that lack MYC and BCL2 or BCL6 translocations (NCCN 2021, Swerdlow 2016).

[0200] PMBCL has distinct clinical, pathological, and molecular characteristics compared with DLBCL and represents approximately 2% to 4% of all patients with NHL (Bhatt 2015, Dabrowska-Iwanicka 2014). Initial treatment for PMBCL is similar to DLBCL (i.e., anthracycline-containing regimens with rituximab with or without involved-field radiotherapy), which is curative in the majority of cases. Chemosensitive patients who relapse may respond to further cytotoxic therapy followed by autologous stem cell transplant (ASCT); nonresponders have limited therapeutic options that lack durability (NCCN 2021).

[0201] Follicular lymphoma 3B (FL3B) is the most aggressive variant of FL, the most common subtype of indolent non-Hodgkin lymphoma. The majority of cases contain a t(14;18) translocation, which juxtaposes BCL2 with the IGH locus. Pathologic grading of FL according to the number of centroblasts is a clinical predictor of outcome. Grade 3 follicular lymphoma is subclassified into FL3A (centrocytes still present) or FL3B (sheets of centroblasts), which have similar survival outcomes but may be managed differently. FL3B is commonly treated according to treatment recommendations for DLBCL (NCCN 2021).

[0202] Treatment options for LBCL include rituximab-based chemotherapy (R-CHOP) and ASCT. Most patients with DLBCL respond well to rituximab-based immunochemotherapy. However, outcome remains poor for patients with r / r DLBCL (1-year and 2-year overall survival [OS] rates of 23% and 16%, respectively). While ASCT confers durable responses, approximately half of DLBCL patients are ineligible for treatment and the majority of ASCT-treated patients will experience disease progression within 3 years (Sehn 2021, Crump 2014, Gisselbrecht 2010). Those patients who do not receive ASCT, either because they are not eligible even after showing response to the second-line chemotherapy or because they do not have responses to the second-line chemotherapy (refractory), have a very poor prognosis with a median overall survival (OS) of 4.4 months. For patients who have relapsed disease within 12 months after ASCT or primary refractory disease, only 26% of them respond to the next line of therapy (third line) with 7% of complete response and median OS of 6.2 months. Patients whose disease progresses after 2 or more prior lines of systemic therapy regimens are unlikely to benefit further from currently available systemic therapy options, unless they have experienced a long disease-free interval. Thus, there is considerable unmet medical need for DLBCL patients who do not respond to first-line immunochemotherapy or to subsequent courses of combination chemotherapy (NCCN 2021).

[0203] To assess the outcomes of patients with r / r LBCL, the multicenter retrospective study SCHOLAR-1 analyzed data from 2 large Phase 3 studies involving 636 patients. Results showed that refractory DLBCL patients have an objective response rate (ORR) of 26%, a complete response (CR) rate of 7%, and a median OS of 6.3 months (Crump 2017, Gisselbrecht 2012, Van Den Neste 2016). Poor outcomes were observed across all refractory subgroups (primary refractory, refractory to second-line or higher therapy, or relapse < 1 year after ASCT) and regardless of disease stage, suggesting that these patients form a single population who lack effective treatment.

[0204] Recently, new therapeutic options have been approved for r / r LBCL. In 2020, FDA granted accelerated approval for tafasitamab in combination with lenalidomide for the treatment of adult patients with r / r DLBCL, NOS, including DLBCL arising from low-grade lymphoma, and who are not eligible for ASCT. Among 71 Page 48 of 173 12403779v1Attorney Docket No. 2017428-0684 subjects with r / r DLBCL treated with tafasitamab plus lenalidomide, 37% achieved a CR and 18% achieved a partial response (PR) (MONJUVI Prescribing Information 2020). The estimated median duration of response (DOR) was 21.7 months. EMA granted approval for polatuzumab vedotin, a CD79b-directed antibody-drug conjugate, in combination with bendamustine and rituximab (BR) for treatment of r / r DLBCL patients who are not candidates for stem cell transplant (SCT). Polatuzumab vedotin plus BR resulted in an ORR of 45% (BR alone: 18%) and a CR rate of 40% (BR alone: 18%) (POLIVY Prescribing Information 2019, Sehn 2018). Pembrolizumab, a humanized monoclonal antibody (mAb) against programmed cell death protein 1 (PD-1), received accelerated approval by FDA for the treatment of PMBCL refractory to treatment or that has relapsed after 2 or more prior lines of therapy (Armand 2019, KEYTRUDA Prescribing Information 2019). The single-arm, multicenter KEYNOTE-170 study of pembrolizumab enrolled 53 patients with a median of 3 prior lines of therapy, and results showed an ORR of 45% (CR: 13%) with the median OS not reached.

[0205] Also recently, autologous CD19-directed chimeric antigen receptor (CAR) T cell therapies (tisagenlecleucel, axicabtagene ciloleucel, and lisocabtagene maraleucel) have been approved as a second-line and / or third-line therapy for patients with r / r LBCL. These therapies offer CR rates ranging from 39% to 60% and median DOR from 9.2 months to 16.7 months. Key toxicities include cytokine release syndrome (CRS) and neurotoxicity with rates of Grade 3 and higher events ranging from 4% to 23% and 11% to 31%, respectively (KYMRIAH® Prescribing Information 2021; YESCARTA® Prescribing Information 2021; BREYANZI® Prescribing Information 2021).

[0206] Three recent randomized trials evaluated CD19-directed CAR T cell therapies as second-line therapy for LBCL (Locke 2022, Bishop 2021, Kamdar 2022). The ZUMA-7 study demonstrated superiority of axicabtagene ciloleucel over standard of care (CR 65% vs 32%, event-free survival (EFS hazard ratio [HR] 0.40, p < 0.001) and TRANSFORM demonstrated superiority of lisocabtagene maraleucel over standard of care (EFS 10.1 months vs 2.3 months, HR 0.35). However, the BELINDA study failed to demonstrate superiority of tisagenlecleucel versus standard of care (EFS HR 1.07, p = 0.69) driven in part by differences in trial design and treatment delays associated with manufacturing.

[0207] The outcome for patients that relapse or are r / r to CD19-directed CAR T cell therapy remains poor, with < 25% responding to subsequent therapies and a median OS of 3.6 months (Spiegel et al., 2021). Therefore, the targeting of alternative antigens represents an important therapeutic strategy for these patients. Research has progressed to investigating CD22-directed CAR T cell therapies to improve the prognosis for those patients that have had a poor response to previous CD19-directed CAR T cell therapy. An ongoing Phase 1 dose escalation*be study reported that a single infusion of autologous CD22-directed CAR T cell therapy achieved CR in 3 patients that had been refractory CD19-directed CAR T cell therapy, with all responses continuing at the time of last follow-up (mean, 7.8 months; range, 6-9.3) (Baird et al., 2021). The same authors presented an abstract of this ongoing study which has since enrolled 38 patients (Frank et al 2023). At dose level (DL) 1 (n = 29), patients were followed for a median of 14.1 months (range, 1.5–38.6 months) and showed an ORR of 66% and a CR rate of 52%. The median progression-free survival (PFS) was 3.0 months (95% confidence interval [CI], 1.6–not reported [NR]) and the median OS was not reached (NR) (95% CI, 8.3–NR). At DL2 (n = 9), the median follow-up was 27.1 months (range, 24.7–33.5 months) and the ORR was 78% with a CR rate of 56%. The median PFS was 2.6 months (95% CI, 1.3–NR) and the median OS was 22.5 months (95% CI, 5.5–NR).

[0208] Despite their approvals, the manufacturing and logistical challenges of autologous CAR T cell therapies make it difficult for these therapies to be widely used (Gajra, 2022). Therefore, considerable unmet Page 49 of 173 12403779v1Attorney Docket No. 2017428-0684 medical need still exists for patients with r / r LBCL, and there is urgent need to develop innovative medicines including more readily available (i.e., off-the-shelf) CAR T cell therapies.

[0209] iNHLs include FL and MZL. FL is the most common indolent form of NHL, accounting for between 10% and 22% of all cases (NCCN 2021, Teras 2016). Although newly diagnosed, low-grade, limited- stage FL is often responsive to a variety of first-line treatments; the disease is characterized by a pattern of relapsing and remitting disease. Transformation from FL to DLBCL (transformed FL [tFL]) occurs at an annual rate of approximately 3% for 15 years and is associated with poor outcomes (Montoto 2007).

[0210] Outcomes in FL are mixed. High-risk patients with ≥ 3 Follicular Lymphoma International Prognostic Index (FLIPI) criteria have a 5-year OS rate of 53%, whereas low-risk patients with ≤ 1 FLIPI criterion have a 5-year survival rate of over 90%. Advanced-stage FL generally requires multiple successive lines of therapy leading to progressively shorter remission periods, chemorefractory disease, transformation to DLBCL, or death due to repeated treatment-related toxicities (Cheah 2018). Thus, regardless of the disease stage at diagnosis, most patients with FL ultimately experience disease relapse, even after a long-term response to first- line therapy, representing a significant unmet need.

[0211] First-line therapies for FL include R-CHOP, R-CVP, bendamustine + CD20 mAb, and lenalidomide + rituximab. High-dose (chemo)therapy / auto-SCT as a consolidative therapy for patients with FL who are in second or third remission is a potential therapeutic option (NCCN 2021). Allogeneic (allo)-SCT may offer improved relapse rates and disease control compared with auto-SCT; however, allo-SCT is associated with higher non-relapse mortality rates and is not a viable treatment option for most patients with FL (Freedman 2018). New targeted agents have been developed to treat patients who progress following systemic chemotherapy and rituximab-containing regimens. The “enhancer of zeste homolog” (EZH)2 histone methyl transferase inhibitor tazemetostat was granted accelerated approval by the FDA for adult subjects with EZH2 mutant r / r FL who have received ≥ 2 prior therapies (or have no satisfactory alternative treatment options) (TAZVERIK 2020). Additionally, phosphatidylinositol 3-kinase inhibitors (i.e., idelalisib, copanlisib, and duvelisib) have been approved for r / r FL after 2 prior therapies.

[0212] CD19-directed CAR T cell therapy has also been utilized in the treatment of r / r FL. In a study of 21 subjects with r / r FL (8 with FL and 13 with tFL) treated with an investigational autologous CD19-directed CAR T therapy, CR rates were 88% and 46% for FL and tFL patients, respectively. All patients with FL who achieved CR remained in remission at a median follow-up of 24 months while the median duration of response for tFL was 10.2 months (Hirayama 2019). YESCARTA has received accelerated approval from the FDA for treatment of adult patients with r / r FL after 2 or more lines of systemic therapy. Approval was based on Study KTEC19105 (ZUMA5), where treatment with YESCARTA among 81 subjects with r / r FL resulted in an ORR of 91% and a CR rate of 60% (YESCARTA 2021). Subjects had at least 9 months of potential follow-up and a median of 3 prior lines of therapy. Among the 74 responding subjects, the median DOR was not estimable with a median follow-up time of 14.5 months.

[0213] These data highlight the need for novel treatment options for patients with r / r FL who have received 2 or more prior lines of therapy and demonstrate the promise of durable responses with CAR T cell therapy.

[0214] MZL is a rare form of NHL originating in the marginal zone of B cells (Kahl 2008). The low incidence rates and heterogeneous presentation render large-scale clinical studies of MZL challenging. First-line Page 50 of 173 12403779v1Attorney Docket No. 2017428-0684 treatment options for MZL include R-CHOP, R-CVP, rituximab, and bendamustine + rituximab. Generally, patients respond to front-line therapy; however, patients with r / r MZL have few therapeutic options, including Bruton tyrosine kinase (BTK) inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, and lenalidomide plus rituximab (NCCN 2021).

[0215] Ibrutinib, a Bruton tyrosine kinase inhibitor (BTKi), is approved for patients with MZL who have received at least 1 prior anti-CD20-based therapy (IMBRUVICA 2022). Accelerated approval of ibrutinib was granted based on a prospective, multicenter, open-label Phase 2 study of 63 patients with MZL that reported an ORR of 46% and CR rate of 3.2%. After a median follow-up time of 19.4 months, the DOR was not reached. Lenalidomide, a thalidomide analogue, is approved for the treatment of previously treated MZL in combination with a rituximab product (REVLIMID 2019). In a pivotal study, among patients with MZL, the ORR by Independent Review Committee assessment was 65% (CR 29%) (Thieblemont 2019). After a median follow-up of 27.9 months, the median progression-free survival (PFS) was 20.2 months.

[0216] Preliminary results with CD19-directed CAR T cell therapy have also demonstrated promising results in r / r MZL. Treatment with YESCARTA among 7 subjects with r / r MZL yielded an ORR of 86% and a CR rate of 71% (Jacobson 2020). To this end, effective modalities to keep patients disease-free are warranted in patients relapsing after 2 lines of therapy and CAR T therapies demonstrate the promise of durable responses. MCL comprises approximately 6% of all newly diagnosed NHL cases.

[0217] Mantle cell lymphoma (MCL) carries the unfavorable characteristics of both indolent and aggressive NHL subtypes due to the incurability of disease with conventional chemotherapy and a generally more aggressive disease course compared to indolent NHL. The MCL International Prognostic Index (MIPI) integrates clinical variables including age, performance status, lactate dehydrogenase, and white blood cell count to stratify patients into low, intermediate, and high-risk categories. High proliferation index (based on KI-67), blastoid / pleomorphic histology, complex karyotype, and TP53 mutations predict poor outcomes for patients (Romancik 2022).

[0218] Initial therapy for MCL often involves intensive, cytarabine-containing induction chemotherapy regimens such as RDHA, HyperCVAD, or NORDIC, followed by consolidation with an autologous stem cell transplant (ASCT) and 3 years of maintenance rituximab. Older patients and those with medical comorbidities may instead receive less intensive induction regimens such as bendamustine plus rituximab, R-CHOP, or lenalidomide plus rituximab. Regardless of initial therapy, most patients will eventually relapse and require multiple lines of subsequent therapies (NCCN 2021).

[0219] Targeted agents such as the BTK inhibitors, bortezomib, lenalidomide, and venetoclax are currently available to treat second-line r / r MCL. Three irreversible BTK inhibitors (ibrutinib, acalabrutinib, and zanubrutinib) are FDA approved for the treatment of r / r MCL. Ibrutinib was the first BTK inhibitor approval based on a pivotal Phase 2 in r / r MCL demonstrating 68% ORR, 21% CR, median DOR 17.5 months, and median 2-year OS of 47% (Wang 2013). Subsequently, more selective, second-generation BTK inhibitors (acalabrutinib and zanubrutinib) were approved. Acalabrutinib’s approval was based on the Phase 2 ACE-LY-2004 trial with 124 r / r MCL patients demonstrating 81% ORR, 40% CR, DOR 26 months and median PFS 19.5 months (Wang 2018). Zanubrutinib’s approval was based on the pivotal BGB-3111-206 and BGB3111-AU-003 trials demonstrating an 84% ORR and 25% to 69% CR (Tam 2021). Lenalidomide is also FDA approved for the treatment of r / r MCL. Page 51 of 173 12403779v1Attorney Docket No. 2017428-0684 When sequenced after bortezomib, lenalidomide monotherapy is associated with an 28% ORR, 7.5% CR, and 16.6 month DOR (Goy 2013).

[0220] Brexucabtagene autoleucel (formerly KTE-X19) is a CD19 autologous CAR T product with a CD28 co-stimulatory domain that is now FDA approved for the treatment of r / r MCL based on the results of the ZUMA-2 trial (Wang 2020). In this study, the ORR was 85% (59% CR) in the intention-to-treat population, with 12-month PFS and OS rates of 61% and 83%, respectively. Notably, responses were observed in patients with high-risk disease characteristics, including pleomorphic / blastoid morphology, TP53 aberrations, and high Ki-67. Lisocabtagene maraleucel (liso-cel; formerly JCAR017) is an autologous CD19-directed CAR T product that contains a 4-1BB signaling domain and is administered by sequential infusions of the CD8+ and CD4+ CAR T cells. Liso-cel is currently FDA approved for the treatment of r / r large B cell lymphomas based on the results of the TRANSCEND NHL 001 trial. While this product is not yet approved for patients with MCL, the Phase 1 portion of the TRANSCEND trial included 32 patients with r / r MCL and reported an ORR of 84% (59% CR) in this population across all dose levels (Abramson 2020).

[0221] Despite the approval of brexucabtagene autoleucel, the product safety profile, manufacturing, and logistical challenges make it difficult for CD19-directed CAR T therapies to be widely used in MCL. Therefore, considerable unmet medical need still exists for patients with r / r MCL, and there is urgent need to develop innovative medicines including more readily available (i.e., off-the-shelf) CAR T cell therapies.

[0222] CLL is the most common leukemia in the USA and among the most common globally. CLL can vary from an indolent disease to very aggressive with potential for transformation to high-grade lymphoma (i.e., Richter’s transformation; NCCN 2022). Previous treatments for CLL included alkylating agents such as chlorambucil and cyclophosphamide and purine nucleosides such as fludarabine, pentostatin, and cladribine. Historically, chemoimmunotherapy regimens like fludarabine, cyclophosphamide and rituximab (FCR), bendamustine plus rituximab (BR), chlorambucil, alemtuzumab, and lenalidomide were the mainstays of CLL treatment in both the front-line and relapsed settings. Over the past decade, BTK inhibitors (ibrutinib, acalabrutinib, zanubrutinib, and pirtobrutinib) BCL2 antagonists (venetoclax), and PI3K inhibitors (idelalisib and duvelisib), given as single agent or in combination with anti-CD20 antibodies (e.g., obinutuzumab), have emerged as new standards of care for front-line CLL. Front-line therapy often leads to durable responses, but despite significant progress, CLL remains incurable. Relapses frequently occur in groups with high-risk features, such as those with TP53 mutation, unmutated immunoglobulin heavy-chain variable (IGHV), deletion 11q or deletion 17p, or presence of complex karyotype. For this reason, chemoimmunotherapies are not recommended for patients with del(17p) / TP53 mutation due to low response rates (Rainone 2022).

[0223] Targeted therapies used in second-line and subsequent settings for CLL include BTK inhibitors, BCL2 antagonists, anti-CD20 antibodies, and PI3K inhibitors. The Phase 3 DUO study of duvelisib, a PI3K delta inhibitor, in 319 patients with r / r CLL demonstrated superior PFS to ofatumumab (13.3 months vs 9.9 months, HR = 0.52, p < 0.001) (Flinn 2018). The Phase 3 IDELA trial in r / r CLL evaluated the PI3K inhibitor idelalisib plus rituximab vs placebo plus rituximab and demonstrated superior efficacy in PFS and OS of the idelalisib-containing arm (Sharman 2019). Few treatment options exist for patients who progress on these targeted therapies and therefore a significant unmet medical need still exists for CLL. Allogeneic stem cell transplantation may provide a chance of cure for some patients but comes with high toxicities and may not be suitable for older CLL patients. Page 52 of 173 12403779v1Attorney Docket No. 2017428-0684

[0224] Recently, CAR T therapies have demonstrated encouraging durable responses for CLL patients. TRANSCEND CLL 004 is the first multicenter study of an CD19-directed CAR T cell therapy (lisocabtagene maraleucel [liso-cel]) for r / r CLL (Siddiqi 2022). Subjects had a median of 4 prior therapies and 83% had high- risk features, including mutated TP53 and del(17p). CRS was reported in 74% of subjects (Grade 3 in 9%), and neurological events in 39% of subjects (Grade 3 / 4 in 22%). Of 22 efficacy-evaluable subjects, 82% and 45% achieved overall and complete responses, respectively. Of 20 minimum residual disease (MRD)-evaluable subjects, 75% and 65% achieved undetectable MRD in blood and marrow, respectively. At a median follow-up of 24 months, the duration of response was not reached and median PFS was 18 months.

[0225] In some embodiments, a subject of the present disclosure has or is suspected of having a B cell malignancy. In some embodiments, a B cell malignancy is selected from the group consisting of: Non- Hodgkin’s Lymphoma (NHL), Chronic Lymphocytic Leukemia (CLL), large B cell lymphoma (LBCL), diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), primary mediastinal B cell lymphoma (PMBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), marginal zone lymphoma (MZL), or small lymphocytic lymphoma (SLL).

[0226] In some embodiments, a subject of the present disclosure has or is suspected of having a CLL with high-risk features, such as those with TP53 mutation, unmutated immunoglobulin heavy-chain variable (IGHV), deletion 11q or deletion 17p, or presence of complex karyotype. In some embodiments, a subject of the present disclosure who has or is suspected of having a CLL with high-risk features is treated with a composition of the present disclosure as a first-line therapy. In some embodiments, a method of the present disclosure comprises detecting one or more biomarkers in a sample from a subject of the present disclosure and identifying the subject as being a high risk CCL patient based on the one or more biomarkers. In some embodiments, the one or more biomarkers are selected from the group consisting of: del(17p), mutated TP53, unmutated immunoglobulin heavy-chain variable (IGHV) gene, and complex karyotype.

[0227] In some embodiments, a subject of the present disclosure who has or is suspected of having multiple myeloma is treated with a composition of the present disclosure as a first-line therapy.

[0228] B cells are thought to have multiple functions that include antigen presentation, cytokine secretion and autoantibody production. There is a plethora of autoantibodies associated with systemic lupus erythematosus (SLE), suggesting continued B cell stimulation and antigen processing in disease. Not all autoantibodies are known to be pathogenic, and autoantibodies are known to appear in advance of clinical disease (Arbuckle 2003). Several studies have highlighted B cell activating factor (BAFF) as having a crucial role in regulating B cell maturation, survival, and function (Vincent 2012). Elevated levels of BAFF have been reported in serum of patients suffering from autoimmune diseases, including SLE, suggesting a role in disease pathogenesis (Renaudineau 2004). Belimumab, a fully humanized mAb which neutralizes BAFF, led to impaired B cell survival and reduction in peripheral B cell numbers in SLE patients (Vincent 2012; Basta 2020). In the Belimumab International Study in Lupus Nephritis 2-year, randomized, placebo-controlled, Phase 3 trial, belimumab plus standard therapies (mycophenolate mofetil [MMF] or cyclophosphamide followed by azathioprine) were administered to adult patients with active lupus nephritis (LN). Compared with placebo, significantly more patients randomized to belimumab achieved a primary efficacy renal response (43% vs.32%; p=0.03) based on reductions in urinary protein to creatine ratio (UPCR) and estimated glomerular filtration rate (eGFR), and a complete renal response (30% vs.20%; p=0.02). Renal-related events or death were also lower Page 53 of 173 12403779v1Attorney Docket No. 2017428-0684 in the belimumab group compared with placebo, with almost 50% lower renal-related events among patients who received belimumab than those who received standard therapies alone (Furie 2020).

[0229] Rituximab, initially approved by the FDA in 1997 for the treatment of r / r NHL, is a mAb that selectively targets the B cell specific surface molecule CD20. The absence of CD20 antigen expression on the cell surface of hematopoietic stem cells, normal plasma cells, or other normal tissues, allows for selective depletion of B cells with rituximab (Fervenza 2008). Murine models have helped establish that CD20+ and circulating B cells are rapidly eliminated by CD20- targeting mAbs, through the reticuloendothelial system. However, B cells in some locations, such as marginal zones, are not depleted by mAbs targeting CD20, and the reliance on the reticuloendothelial system for B cell depletion is problematic in SLE where it is known to be compromised (Gong 2005).

[0230] Two large, randomized, placebo-controlled, Phase 2 studies in non-renal lupus (EXPLORER) and LN (LUNAR) both failed to meet their primary endpoints after treatment with rituximab, although both trials demonstrated partial responses in selected patients (Merrill 2010; Rovin 2012). Complete peripheral depletion of B cells with rituximab was not observed in all participants, and even in participants where complete peripheral depletion of B cells was observed, <50% achieved complete response (Gomez 2018).

[0231] As observed in murine models, the absence of a positive correlation between adequate peripheral depletion of B cells and complete response in the trial participants suggests that autoreactive B cells may persist in protected microenvironments like the lymphoid structures, kidney tubulointerstitial and bone marrow and therefore prolonged exposure to rituximab (>52 weeks) is needed (Gong 2005; Ahuja 2007; Ahuja 2011; Bekar 2010). Combination therapies involving rituximab and belimumab (a BAFF inhibitor) have been shown to reduce the re-emergence of autoreactive B cells following B cell depletion, enhance negative selection of autoreactive B cells, and lower anti-dsDNA antibody levels (Atisha-Fregoso 2021; van Schaik 2022). While combination treatments are effective in some patients, certain severe forms of SLE remain resistant to treatment.

[0232] Currently, there is no standard of care (SoC) treatment for achieving drug-free remission of SLE; therefore, patients often require life-long therapy. While a combination approach using antimalarials (hydroxychloroquine), systemic steroids, and conventional immunosuppressant medicines (azathioprine, MMF, and cyclophosphamide) are first-line options for SLE treatment, a significant proportion of patients continue to have high disease activity and recurrent relapses (Ginzler 2005; Rahman 2008; Appel 2009). Despite the apparent effectiveness of these regimens, complete renal remission in 30–50% of patients within the first year, between 20–35% of patients experience renal relapse on maintenance therapy over a 3–6-year follow-up period, with one study reporting that only 38% of patients were able to maintain a complete renal response over a 5- year period. A lack of response to therapy after 1 year of treatment was significantly associated with increased mortality and chronic kidney disease risk (p<0.005) (Anders 2015) and cyclophosphamide carries a secondary malignancy risk, in addition to increased risk of infertility with cumulative exposure (Ginzler 2005).

[0233] After the results of the LUNAR trial demonstrated no benefit of rituximab in complete renal response and a 15% improvement in partial renal response, a retrospective investigation was undertaken evaluating peripheral blood B cell depletion and occurrence of complete renal response. The data demonstrated that deeper B cell depletion was associated with improved complete renal response, and that poor peripheral blood B cell depletion was associated with non-response. This observation (Gomez 2018), coupled with data demonstrating that obinutuzumab (a type II mAb with greater antibody dependent cellular cytotoxicity through Page 54 of 173 12403779v1Attorney Docket No. 2017428-0684 glycoengineering) in CLL was superior to rituximab in progression-free survival and minimal residual disease negativity in a head-to-head trial of CLL (Goede 2014) prompted the initiation of a Phase 2 trial of Obinutuzumab in LN. The hypothesis driving this trial was that deeper B cell depletion should increase complete renal response in LN patients. The Phase 2 results were published in 2021 (Furie 2022) and described a 12% improvement in complete renal response and 21% partial renal response at 52 weeks in the obinutuzumab-treated patients vs controls. The Phase 3 trial has completed enrolment and results are expected in 2024.

[0234] The current SoC treatment for anti-neutrophilic cytoplasmic autoantibody (ANCA) associated vasculitis (AAV) involves a combination of high dose glucocorticoids with either cyclophosphamide or rituximab, with evidence suggesting that the combination of rituximab and glucocorticoids is superior for remission induction in severe relapsing AAV compared with cyclophosphamide treatment alone (Stone 2010). Cyclophosphamide therapy is designed to target proliferating cells, resulting in blunted expansion of pathogenic clones, while memory B and T cells are unaffected. However, due to the dose-limiting AEs induced by cyclophosphamide, coupled with severe toxicity, complete depletion of pathogenic clones is rare, resulting in treatment failure and risk of chronic relapse. Cyclophosphamide is also associated with leukopenia, cancer, and ovarian failure (Mukhtyar 2008). Severe infections are also associated with prolonged cyclophosphamide exposure and are predominant causes of early death in patients (Jones 2010).

[0235] Results from the RAVE trial found that rituximab was non-inferior to cyclophosphamide for remission of disease defined as Birmingham Vasculitis Activity Scale (BVAS) / WG=0, without the use of prednisone, at 6 months (64% in rituximab group vs 53% in cyclophosphamide group) (Stone 2010). In addition, rituximab was superior to cyclophosphamide for induction of remission for relapsing disease (67% in rituximab group, 42% in cyclophosphamide group). Finally, there were no reported significant differences in rates of adverse events between the two treatments. Despite rituximab showing superiority against cyclophosphamide in reducing relapse rates, remission rates are still high in rituximab-treated patients, especially in patients with a history of relapse. The RITAZAREM trial demonstrated that repeated dosing of rituximab following remission (defined as BVAS / WG =0) in patients was superior compared to azathioprine at preventing relapse (HR 0.41; 95% CI 0.27 to 0.61, p<0.001) (Smith 2023). The MAINRITSAN trials demonstrated that prolonged rituximab treatment was an effective regimen in sustaining remission (defined as BVAS =0) in patients, however, for some patients after 2 years of maintenance, relapse risk remains high and extended therapy is advised (Tieu 2020; Charles 2018; Charles 2020; Guillevin 2014). Despite promising results with rituximab, there remains an unmet treatment need for this group of patients as highlighted in a recent study investigating the long-term outcomes and prognostic factors for survival of patients with AAV (Sanchez 2023). The study concluded that patients with AAV had an increased risk of mortality when compared to the general population, with treatment complications and organ damage being the main causes of limited survival and infection the leading cause of mortality.

[0236] Improved treatment options are needed as conventional therapies are not only associated with treatment failures and relapses, but also high levels of toxicity which result in severe morbidity and lethal adverse effects (Jayne 2003). A recent randomized controlled trial (ADVOCATE) investigated the C5a receptor inhibitor, avacopan, for the treatment of AAV (Jayne 2021). Avacopan is a C5a receptor antagonist that selectively blocks the effects of C5a preventing neutrophil chemoattraction and activation and has demonstrated beneficial effects on vasculitis in Phase 2 clinical trials (Jayne 2017; Merkel 2020). All patients were given either cyclophosphamide or rituximab and then assigned into two groups: one received avacopan and the other prednisone. Results showed avacopan was non-inferior but not superior to prednisone in inducing remission Page 55 of 173 12403779v1Attorney Docket No. 2017428-0684 defined as BVAS=0 of vasculitis at 26 weeks and was superior to prednisone at 52 weeks in patients who received rituximab or cyclophosphamide.

[0237] In some embodiments, a subject of the present disclosure has or is suspected of having a disease, disorder, or condition that is relapsed and / or refractory. In some embodiments, a B cell malignancy is relapsed and / or refractory. In some embodiments, a B cell malignancy is relapsed and / or refractory NHL. In some embodiments, a B cell malignancy is relapsed and / or refractory CLL. In some embodiments, a B cell malignancy is relapsed and / or refractory LBCL. In some embodiments, the B cell malignancy is relapsed and / or refractory after at least 2 regimens of therapy. In some embodiments, a B cell malignancy is relapsed and / or refractory after autologous stem cell transplant (ASCT). In some embodiments, a B cell malignancy is relapsed and / or refractory, wherein relapsed and / or refractory comprises one or more of: no response to first line therapy, progressive disease as best response to most recent therapy regimen, stable disease as best response to most recent therapy with duration no longer than 6 months from last dose of therapy, disease progression or relapse within 12 months of ASCT, no response to or relapse after last line of therapy post-ASCT, and / or no response to or relapse after treatment with a first CD19 CAR T therapy. In some embodiments, a subject of the present disclosure has or is suspected of having relapsed and / or refractory CLL and has already been treated with at least 2 regimens of therapy. In some embodiments, a subject of the present disclosure has or is suspected of having relapsed and / or refractory multiple myeloma and has already been treated with at least 1 regimen of therapy. In some embodiments, a subject of the present disclosure has or is suspected of having relapsed and / or refractory LBCL and is transplant ineligible. In some embodiments, a subject of the present disclosure has already been treated with a bispecific antibody. In some embodiments, a bispecific antibody is selected from the group consisting of: mosunetuzumab (CD20 x CD3 mAb), glofitamab (CD20(2) x CD3), odronextamab (CD20 x CD3 mAb), and epcoritamab (CD20 x CD3 mAb). In some embodiments, a B cell mediated disorder is relapsed and / or refractory. In some embodiments, a B cell mediated disorder is relapsed and / or refractory following a first line therapy comprising mycophenolate mofetil (MMF), cyclophosphamide, or a combination thereof. In some embodiments, a B cell mediated disorder is relapsed and / or refractory following a second line therapy comprising rituximab, belimumab, voclosporin, obinutuzumab, or a combination thereof. In some embodiments, a B cell mediated disorder is relapsed and / or refractory following at least two disease- modifying therapies selected from the group consisting of: MMF, azathioprine, cyclophosphamide, methotrexate, rituximab, belimumab, intravenous immunoglobulin (IVIg), and abatacept. In some embodiments, a subject does not achieve a complete renal response to the second line therapy within 6 months of initiation of the second line therapy. In some embodiments, a subject has disease activity despite maintenance on maximally tolerated doses of drugs that block the renin-angiotensin system (RAS).

[0238] In some embodiments, a subject of the present disclosure is 18-85 years of age. In some embodiments, a subject of the present disclosure is 18-80 years of age. In some embodiments, a subject of the present disclosure is 18-75 years of age. 4. Exclusion criteria

[0239] In some embodiments, a subject of the present disclosure cannot or should not be treated with a pharmaceutical composition of the present disclosure if they meet certain criteria (exclusion criteria). In some embodiments, a subject of the present disclosure has not received a prior CD22-directed therapy. In some embodiments, a prior CD22-directed therapy is or comprises a CD22-directed CAR T cell treatment. In some embodiments, a prior CD22-directed therapy is or comprises a CD22-directed antibody treatment. In some Page 56 of 173 12403779v1Attorney Docket No. 2017428-0684 embodiments, a prior CD22-directed therapy is or comprises a CD22-directed CAR NK cell treatment. A CD47- directed therapy can includes a binder of CD47 (e.g., an anti-CD47 antibody) that specifically binds to the CD47 and interferes with the interaction of CD47 and SIRPα. Examples of binders that specifically bind CD47 include magrolimab, Trillium (TTI)-621, ALX148, AO-176, lemzoparlimab, IBI188, SRF231, CC-90002, and AK117. A CD47-directed therapy can also include a protein and / or molecule that interferes with the interaction of CD47 and SIRPα by binding SIRPα, such as ADU-1805 and BI 765063. In some embodiments, a subject has not received a CD47-directed therapy within 1 year of being administered the composition comprising the population of engineered hypoimmunogenic T cells. In some embodiments, a subject has not received a CD47-directed therapy within about 1 year, about 11 months, about 10 months, about 9 months, about 8 months, about 7 months, about 6 months, about 5 months, about 4 months, about 3 months, about 2 months, or about 1 month of being administered the composition comprising the population of engineered hypoimmunogenic T cells. In some embodiments, a subject of the present disclosure having or suspected of having anti-CD47 antibodies cannot or should not be treated with a pharmaceutical composition of the present disclosure until anti-CD47 antibody levels are reduced. In some embodiments, a subject of the present disclosure having or suspected of having anti-CD47 antibodies cannot or should not be treated with a pharmaceutical composition of the present disclosure until a period of time known to be associated with reduced (i.e., cleared) anti-CD47 antibody levels has passed. For example, the period of time may be 1 year, about 11 months, about 10 months, about 9 months, about 8 months, about 7 months, about 6 months, about 5 months, about 4 months, about 3 months, about 2 months, or about 1 month. As another example, the period of time may be 5, 6, 7, 8, 9, or 10 half-lives of the CD47-directed therapy. In some embodiments, a subject of the present disclosure having or suspected of having anti-CD47 antibodies cannot or should not be treated with a pharmaceutical composition of the present disclosure until testing of anti-CD47 antibody levels indicates reduced levels. In some embodiments, a subject of the present disclosure having or suspected of having anti-CD47 antibodies cannot or should not be treated with a pharmaceutical composition of the present disclosure until anti-CD47 antibody levels are at undetectable levels. Any method known in the art for detecting antibodies in a sample from a patient can be used. For example, antibodies can be detected in a sample (e.g., serum or whole blood) using an enzyme-linked immunospot (ELISpot) assay, ELISA, immunofluorescence, flow cytometry, Western blot, or any other methodology of determining the antibody titer of a specific antibody. In some embodiments, a subject of the present disclosure subject does not have a history of primary central nervous system (CNS) lymphoma or CNS metastases. In some embodiments, a subject of the present disclosure does not have a history of Richter’s transformation of chronic leukemic lymphoma, small lymphocytic lymphoma, or lymphoplasmacytic lymphoma. In some embodiments, a subject of the present disclosure does not have a fungal, bacterial, viral, or other infection that is uncontrolled or requiring IV antimicrobials for management. In some embodiments, a subject of the present disclosure has not been hospitalized for treatment of an infection within 28 days of baseline of > 2 times within 1 year of administering the composition to the subject. In some embodiments, a subject of the present disclosure does not have acute symptoms of COVID-19 infection. In some embodiments, a subject of the present disclosure does not have a known history of infection with HIV and / or hepatitis B (HBsAg positive) and / or hepatitis C virus (anti-HCV positive). In some embodiments, a subject of the present disclosure does not have an active autoimmune disease and / or any other diseases requiring immunosuppressive therapy or corticosteroid therapy. In some embodiments, a subject of the present disclosure does not have primary immunodeficiency or history of concomitant genetic syndrome associated with bone marrow failure (e.g., Fanconi anemia, Kostmann syndrome, or Shwachman-Diamond). In some embodiments, a subject of the present disclosure has not been treated with Page 57 of 173 12403779v1Attorney Docket No. 2017428-0684 a systemic cancer therapy within 14 days of administration of the composition. In some embodiments, a subject of the present disclosure has not been treated with any investigational product within 28 days of administration of the composition. In some embodiments, a subject of the present disclosure has not been treated with radiotherapy within 14 days of administration of the composition. In some embodiments, a subject of the present disclosure has not been treated with major surgery within 28 days of administration of the composition. In some embodiments, a subject of the present disclosure has not been treated with autologous hematopoietic stem cell transplant (HSCT) within 6 weeks of administration of the composition. In some embodiments, a subject of the present disclosure has not had myocardial infarction, cardiac angioplasty or stenting, cardiac arrhythmia requiring medication, unstable angina, New York Heart Association Class II or greater congestive heart failure, cardiac atrial or ventricular lymphoma involvement or clinically significant cardiac disease within one year of administration of the composition. In some embodiments, a subject of the present disclosure has not had an ejection fraction ≤ 50%, arrythmias and any structural heart disease (including clinically significant pericardial effusion), as assessed by echocardiogram or multiple gated acquisition scan performed within 1 month of administration of the composition. In some embodiments, a subject of the present disclosure does not have a history of non-line associated, clinically significant deep-vein thrombosis (i.e., proximal deep vein thrombosis [DVT]) or pulmonary embolism requiring therapeutic anticoagulation within 6 months of administration of the composition. In some embodiments, a subject of the present disclosure does not have a history or presence of CNS disorder (e.g., seizure disorder, cerebrovascular ischemia / hemorrhage, dementia, cerebellar disease, or any autoimmune disease with CNS involvement) within 12 months of administration of the composition. In some embodiments, a subject of the present disclosure does not have clinically significant ascites. In some embodiments, a subject of the present disclosure does not have a history of organ transplantation and / or is not awaiting organ transplantation. In some embodiments, a subject of the present disclosure does not have an indwelling line or drain. In some embodiments, a subject of the present disclosure has not received a live vaccine within 6 weeks of administration of the composition. In some embodiments, a subject of the present disclosure does not have any unresolved toxicity greater than Grade 1 from a previous anticancer therapy. In some embodiments, a subject of the present disclosure does not have a history of additional malignancy other than nonmelanoma skin cancer or carcinoma in situ. In some embodiments, a subject of the present disclosure does not have a severe immediate hypersensitivity to any of the components of a pharmaceutical composition of the present disclosure. In some embodiments, a subject of the present disclosure does not have an admission or evidence of illicit drug use, drug abuse, or alcohol abuse. In some embodiments, a subject of the present disclosure does not exhibit clinical evidence of significant unstable or uncontrolled acute or chronic disease not due to the B cell mediated disorder. In some embodiments, a significant unstable or uncontrolled acute or chronic disease comprises cardiovascular, pulmonary, hematologic, gastrointestinal, hepatic, renal, neurological, malignancy, or infectious disease. In some embodiments, a subject of the present disclosure does not have a history of an anaphylactic reaction to parenteral administration of contrast agents, human or murine proteins, or monoclonal antibodies. In some embodiments, a subject of the present disclosure does not have a history of cancer, or the subject has been disease-free from a basal carcinoma of the skin, cervix, bladder, prostate or breast for at least five years prior to administration of the composition. In some embodiments, a cancer comprises a hematologic malignancy, a solid tumor, or a carcinoma in situ. In some embodiments, a subject of the present disclosure is administered physiologic replacement glucocorticoids, topical glucocorticoids, or inhaled glucocorticoids. In some embodiments, a subject of the present disclosure is not administered glucocorticoids above physiologic replacement. In some Page 58 of 173 12403779v1Attorney Docket No. 2017428-0684 embodiments, physiologic replacement glucocorticoids comprises 5 mg prednisone daily or equivalent. In some embodiments, a subject of the present disclosure has not received treatment with Leflunomide within 28 prior to administration of the composition. In some embodiments, a subject of the present disclosure does not have an immunoglobulin A (IgA) deficiency (IgA level < 10mg / dL). In some embodiments, a subject of the present disclosure does not have a grade ≥ 3 laboratory abnormality, except those associated with the B cell mediated disorder. In some embodiments, a subject of the present disclosure does not have a B cell mediated disorder related flare that needs immediate treatment. In some embodiments, a subject of the present disclosure does not have a CNS lupus manifestation, a history of a CNS disorder, or a presence of a CNS disorder. In some embodiments, a CNS disorder is or comprises primary seizure disorder, cerebrovascular ischemia / hemorrhage, dementia, or cerebellar disease. In some embodiments, a subject of the present disclosure has not been diagnosed with anti-phospholipid antibody syndrome that is actively treated. In some embodiments, a subject of the present disclosure has not been diagnosed with EGPA as defined by the Chapel Hill Consensus Conference criteria. In some embodiments, a subject of the present disclosure does not have an alveolar hemorrhage within 4 weeks prior to administration of the composition. In some embodiments, a subject of the present disclosure has not been diagnosed with a cerebrovascular accident caused by vasculitis or pachymeningitis.

[0240] In some embodiments, a subject of the present disclosure should not take corticosteroid therapy at a pharmacologic dose (≥ 5 mg / day of prednisone or equivalent doses of other corticosteroids) and other immunosuppressive drugs within 5 days of administration of a lymphodepletion regimen of the present disclosure and / or administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure.

[0241] In some embodiments, a subject of the present disclosure should not take corticosteroids and / or other immunosuppressive drugs in the 3 months after administration of hypoimmune allogenic CD22- directed CAR T cells of the present disclosure. In some embodiments, a subject of the present disclosure should not take nonsteroidal anti-inflammatory agents in the 3 months after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, a subject of the present disclosure should not take therapeutic doses of systemic anticoagulants, such as unfractionated heparin and low-molecular weight heparin after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, a subject of the present disclosure does not receive treatments for underlying disease, such as surgery, chemotherapy, immunotherapy, targeted agents, radiation, and high-dose corticosteroids (other than those allowed herein) and other investigational agents after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. 5. Lymphodepletion regimen

[0242] Non-limiting examples of an immunosuppressive and / or immunomodulatory agent (such as, but not limited to a lymphodepletion agent) include cyclosporine, azathioprine, mycophenolic acid, mycophenolate mofetil, corticosteroids such as prednisone, methotrexate, gold salts, sulfasalazine, antimalarials, brequinar, leflunomide, mizoribine, 15-deoxyspergualine, 6-mercaptopurine, cyclophosphamide, rapamycin, tacrolimus (FK-506), OKT3, anti-thymocyte globulin, thymopentin, thymosin-α and similar agents. In some embodiments, the immunosuppressive and / or immunomodulatory agent is selected from a group of immunosuppressive antibodies consisting of antibodies binding to p75 of the IL-2 receptor, antibodies binding to, for instance, MHC, CD2, CD3, CD4, CD7, CD28, B7, CD40, CD45, IFN-gamma, TNF-alpha, IL-4, IL-5, IL-6R, IL-6, IGF, IGFR1, IL-7, IL-8, IL-10, CD11a, or CD58, and antibodies binding to any of their ligands. In some embodiments, such an immunosuppressive and / or immunomodulatory agent may be selected from soluble IL- Page 59 of 173 12403779v1Attorney Docket No. 2017428-0684 15R, IL-10, B7 molecules (e.g., B7-1, B7-2, variants thereof, and fragments thereof), ICOS, and OX40, an inhibitor of a negative T cell regulator (such as an antibody against CTLA-4) and similar agents.

[0243] In some embodiments, where an immunosuppressive and / or immunomodulatory agent is administered to the patient before or after the administration of the cells, the administration is at a lower dosage than would be required for cells with MHC I and / or MHC II expression, TCR expression and without exogenous expression of CD47. In some embodiments, where an immunosuppressive and / or immunomodulatory agent is administered to the patient before or after the first administration of the cells, the administration is at a lower dosage than would be required for cells with MHC I and MHC II expression, TCR expression and without exogenous expression of CD47.

[0244] In some embodiments, a subject of the present disclosure is administered a lymphodepleting chemotherapy regimen, wherein administering the lymphodepleting chemotherapy regimen occurs prior to administering a pharmaceutical composition of the present disclosure. In some embodiments, a lymphodepleting therapy is or comprises cyclophosphamide and fludarabine. In some embodiments, cyclophosphamide is administered at a dose of 500 mg / m2. In some embodiments, cyclophosphamide is administered at a dose of 500 mg / m2daily for 3 days. In some embodiments, cyclophosphamide is administered at a dose of 1000 mg / m2. In some embodiments, cyclophosphamide is administered at a dose of 1000 mg / m2one, 3 days prior to administration of a composition of the present disclosure. In some embodiments, fludarabine is administered at a dose of 30 mg / m2. In some embodiments, fludarabine is administered at a dose of 25 mg / m2. In some embodiments, a subject of the present disclosure has a moderate impairment of renal function, and fludarabine is administered at a dose of about 20 mg / m2with monitoring. In some embodiments, fludarabine is administered at a dose of 30 mg / m2daily for 3 days. In some embodiments, cyclophosphamide and fludarabine are administered concurrently. In some embodiments, a lymphodepleting chemotherapy regimen is administered intravenously. In some embodiments, a pharmaceutical composition of the present disclosure is administered to a subject 2-7 days after completion of the lymphodepleting chemotherapy regimen.

[0245] In some embodiments, a subject of the present disclosure has not received a live vaccine within 28 days of administration of the lymphodepleting chemotherapy.

[0246] In some embodiments, a subject of the present disclosure is weaned to discontinuation of a severe autoimmune therapy over the course of no more than 21 days prior to administration of a composition of the present disclosure to the subject. In some embodiments, a subject is weaned to 0 mg glucocorticoids. In some embodiments, a subject is weaned to a physiologic equivalent range of ≤ 5 mg / day of prednisone.

[0247] In some embodiments, a subject is not administered a lymphodepleting chemotherapy regimen prior to administration of a pharmaceutical composition of the present disclosure. In some embodiments, the following lymphodepletion protocol is followed for a subject of the present disclosure. On Days -5 through -3, before the infusion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure, subjects receive a non-myeloablative lymphodepletion (LD) regimen consisting of cyclophosphamide and fludarabine to induce lymphocyte depletion and create an optimal environment for the expansion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure in vivo. Hypoimmune allogenic CD22-directed CAR T cells of the present disclosure should be administered 3 days (range 2-7 days) after completion of LD chemotherapy.

[0248] In some embodiments, subjects of the present disclosure receive the following 3-day chemotherapy regimen per the order identified as follows: 1) IV hydration with 1 L of 0.9% sodium chloride Page 60 of 173 12403779v1Attorney Docket No. 2017428-0684 (NaCl) given before cyclophosphamide on the day of infusion; 2) Cyclophosphamide 500 mg / m2IV over approximately 60 minutes on Days -5, -4, and -3; 3) Fludarabine 30 mg / m2 IV over approximately 30 minutes on Days -5, -4, and -3 and subjects with moderate impairment of renal function (creatinine clearance 50-70 mL / min / 1.73 m2) should have their fludarabine dose reduced by 20% and be monitored closely; 4) an additional 1 L of 0.9% NaCl at the completion of the fludarabine infusion; and 5) add Mesna (sodium 2- mercaptoethanesulfonate; a detoxifying agent used to inhibit the hemorrhagic cystitis induced by the chemotherapy) per institutional guidelines. Subjects at risk for tumor lysis syndrome (TLS) should receive TLS prophylaxis prior to LD chemotherapy according to institutional guidelines.

[0249] Administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure can increase the risk of high-grade and fatal toxicities in subjects with ongoing infection and / or inflammation. In some embodiments, if a subject of the present disclosure has: a temperature > 38⁰C within 72 hours before LD chemotherapy; CRP ≥ 100 mg / L any time between enrollment and the start of LD chemotherapy; and / or a white blood cell (WBC) count or WBC differential concerning for infectious process between enrollment and the start of LD chemotherapy (e.g., WBC > 20,000 cells / μL, rapidly increasing WBC, or WBC differential with a high percentage of segments / bands), the cause of the infection should be identified and treated before administration of an LD regimen of the present disclosure.

[0250] In some embodiments, if any screening assessments or procedures are repeated between confirmation of eligibility and the start of LD chemotherapy and the results are outside of the eligibility criteria provided herein, any suspected / identified infectious / inflammatory process condition must resolve before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, a complete medical history and physical examination of a subject of the present disclosure including the head, eyes, ears, nose, and throat (HEENT) and cardiac, vascular, respiratory, gastrointestinal, integumentary, and neurological systems does not reveal evidence of infection / inflammation before administration of an LD regimen of the present disclosure. In some embodiments, a subject of the present disclosure has not received systemic antimicrobials for the treatment of a known or suspected infection within the 48 hours before an LD regimen of the present disclosure (prophylactic use of antimicrobials is allowed). In some embodiments, a treatment course of any antimicrobials given for a known or suspected antecedent infection should be completed before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, if a subject of the present disclosure is confirmed to have an infectious process for which antimicrobials are not available (e.g., viral pneumonia), the infection must be clinically resolved before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, the most recently collected blood, urine, or other body fluid cultures must show no growth for at least 48 hours, and any other infectious workup performed (e.g., bacterial, PCR, or stool or imaging studies) is negative before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, if there is clinical suspicion of an infection for which cultures are unlikely to be positive within 48 hours (e.g., fungal infection), then adequate time is allowed to allow the cultures to become positive before a subject of the present disclosure is administered an LD regimen of the present disclosure. 6. Assessment of Subjects

[0251] In some embodiments, subjects of the present disclosure undergo safety assessments that include the collection of adverse events (AEs) and serious adverse events (SAEs), clinical laboratory tests, Page 61 of 173 12403779v1Attorney Docket No. 2017428-0684 imaging, physical and neurologic examinations, vital signs, cardiac function (e.g., electrocardiogram [ECG]), and ECOG performance status, as described herein. In some embodiments, subjects of the present disclosure asked about any AEs that might have occurred since a previous evaluation. In some embodiments, serious adverse events (SAEs), treatment-related adverse events (TRAEs), treatment-emergent adverse events (TEAEs), targeted adverse events (TAEs), adverse events of special interest (AESIs), and / or dose-limiting toxicities (DLTs) are determined for a subject as part of an evaluation.

[0252] In some embodiments, subjects of the present disclosure undergo safety assessments that include clinical laboratory tests. Exemplary clinical laboratory tests include those found in TABLE 10A, TABLE 10B or TABLE 11 of the present disclosure. In some embodiments, subjects of the present disclosure undergo safety assessments that include physical examination. In some embodiments, a physical examination comprises measurement of, assessment of, or performance of one or more of: height, weight, vital signs (e.g., systolic / diastolic blood pressure, heart rate, respiratory rate, oxygen saturation, temperature), neurological status (e.g., immune effector cell-associated encephalopathy (ICE) score), Eastern Cooperative Oncology Group (ECOG) score, cardiac function, echocardiogram (ECHO), electrocardiogram (ECG), brain Magnetic Resonance Imaging (MRI), a lumbar puncture, and information about concomitant medications.

[0253] In some embodiments, subjects of the present disclosure are assessed for the presence of replication competent lentivirus (RCL) in peripheral blood mononuclear cells (PBMCs). In some embodiments, subjects of the present disclosure are assessed for abnormal T cell proliferation. In some embodiments, subjects of the present disclosure are assessed for the presence of anti-CD22-directed CAR antibodies in the subjects’ serum. In some embodiments, serum from subjects of the present disclosure is assessed for cytokine profiling.

[0254] In some embodiments, subjects of the present disclosure are assessed for disease progression and / or disease response. In some embodiments, subjects of the present disclosure with NHL will be assessed based on Lugano classification criteria. In some embodiments, subjects of the present disclosure with CLL will be assessed based on the International Workshop on Chronic Lymphocytic Leukemia (iwCLL) criteria. In some embodiments, disease assessment in subjects of the present disclosure comprises imaging. In some embodiments, imaging comprises positron emission tomography (PET)-computed tomography (CT) and / or CT scans. In some embodiments, subjects of the present disclosure undergo PET-CT and / or CT scans within 24 months of administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, disease assessment in subjects of the present disclosure comprises bone marrow aspirate and / or biopsy.

[0255] In some embodiments, PBMC samples from subjects of the present disclosure are assessed for cellular kinetics of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, cellular kinetics are evaluated by droplet digital polymerase chain reaction (ddPCR) and reported as a level of hypoimmune allogenic CD22-directed CAR T cells per μg of genomic DNA (gDNA). In some embodiments, the derived parameters of Cmax, Tmax, t1 / 2, Clast, Tlast, and AUC are determined for hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, the number of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure in a subject’s blood will be assessed via flow cytometry. In some embodiments, cellular kinetics as described herein are used to determine dose- exposure relationships and / or correlation of persistence and expansion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure with safety and efficacy parameters. Page 62 of 173 12403779v1Attorney Docket No. 2017428-0684

[0256] In some embodiments, blood / serum samples from subjects of the present disclosure undergo pharmacodynamic assessments. In some embodiments, pharmacodynamic assessments comprise monitoring of levels of analytes in blood / serum over time. In some embodiments, analytes include one or more of: homeostatic / proliferative cytokines (e.g., interleukin (IL)-2, IL-7, IL-15, and granulocytecolony-stimulating factor (G-CSF)), inflammatory / immune‐modulating cytokines (e.g., interferon-gamma (IFN-γ), IFN-α, IL-1β, IL-1Ra, IL- 4, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-17A, granulocyte macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-α), macrophage inflammatory protein-1 Alpha (MIP-1α), and MIP-1β), correlates of acute phase response (e.g., C-reactive protein (CRP) and ferritin), chemokines (e.g., IL-8, C-X-C motif chemokine ligand-10 (CXCL-10), and monocyte chemotactic protein-1 (MCP-1)), and associated response related with CD22-directed CAR activity (e.g., circulating CD22+ B cell and lactate dehydrogenase (LDH)).

[0257] In some embodiments, subjects of the present disclosure are assessed for minimal residual disease (MRD). In some embodiments, MRD is determined using a next-generation sequencing (NGS) MRD assay to measure ctDNA levels before and after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, subjects of the present disclosure are assessed for immune responses against the administered hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, immune responses hypoimmune allogenic CD22-directed CAR T cells of the present disclosure are or comprise anti-hypoimmune allogenic CD22-directed CAR T cell antibodies, antibody- dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), immune activation against hypoimmune allogenic CD22-directed CAR T cells of the present disclosure, NK cell-mediated killing of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure.

[0258] In some embodiments, methods of the present disclosure comprise measuring MRD in a subject to monitor treatment response or relapse of disease. In some embodiments, MRD refers to a small number of disease cells that remain in the subject during or after treatment when the subject is in remission. In remission, the subject may no longer display obvious symptoms or signs of disease, but MRD cells may remain and cause relapse of the disease. Testing for MRD may be useful for determining whether treatment has eradicated disease cells or whether traces remain, comparing efficacy of different treatments, monitoring patient remission status as well as detecting recurrence of the disease, and adjusting treatments accordingly. The number of MRD cells in a sample from the subject may be as low as one disease cell in a million normal cells.

[0259] In some embodiments, MRD can be measured using various methods as described in, but not limited to, US20220251654A1, US20170335391A1, US20210238694A1, and US20200370129A1.

[0260] In some embodiments, measuring MRD comprises collecting two or more biological samples from a subject at different time points (e.g. prior to, concurrent with, or after administration of hypoimmune allogenic CD22-directed CAR T cells). In some embodiments, a multiplex amplification reaction is performed on nucleic acids isolated from the biological samples to generate a set of amplicons, wherein each of the set of amplicons comprises recombined V(D)J gene segments at a gene locus of interest and / or wherein each of the set of amplicons comprises gene segments at one or more cancer specific gene loci of interest, and sequencing the set of amplicons, wherein sequences of the recombined V(D)J gene segments and / or cancer specific gene segments are indicative of presence of an immune cell and / or cancer in the biological sample.

[0261] In some embodiments, measuring MRD comprises assigning a pair of first and second polypeptides that form a TCR (TCRA or TCRB) or immunoglobulin (e.g., Ig heavy chain, or κ or λ light chain) Page 63 of 173 12403779v1Attorney Docket No. 2017428-0684 heterodimer to a single biological source sample among the two or more biological samples. In some embodiments, the first rearranged nucleic acid sequences encoding the first polypeptides of the TCR or Ig heterodimers present in the source biological sample is determined for each of the plurality of biological samples and the first rearranged nucleic acid sequences are assigned to the source biological sample. The plurality of source biological samples are then pooled to form a combined population of cell and a plurality of cognate pairs of first and second rearranged nucleic acid sequences encoding first and second polypeptides of the TCR or Ig heterodimers is determined from the combined population of cells. The first rearranged nucleic acid sequences determined in each of the source samples are compared to the first rearranged nucleic acid sequences determined from the plurality of cognate pairs of rearranged nucleic acid sequences to assign each first rearranged nucleic acid sequence present in the combined population to a single biological source sample. For each first rearranged nucleic acid sequence assigned to a single biological source sample, the cognate second rearranged nucleic acid sequence of the cognate pair is assigned to the same single biological source sample.

[0262] In another embodiment, MRD is measured by determining a first rearranged nucleic acid sequence encoding the first polypeptide of the TCR or Ig heterodimer present in the biological source sample. In some embodiments, for each biological source sample, rearranged nucleic acid molecules extracted from the biological source sample are amplified in a single multiplex polymerase chain reaction (PCR) using a plurality of V-segment primers and a plurality of J-segment primers to produce a plurality of rearranged nucleic acid amplicons. The plurality of rearranged nucleic acid amplicons are sequenced to determine sequences of the first rearranged nucleic acid sequences in each biological source sample.

[0263] In some embodiments, MRD is measured by using a “bait set” of probes for hybridization capture. The bait set comprises a plurality of different oligonucleotide-containing probes, where each of the oligonucleotide-containing probes comprises a sequence of at least 30 bases in length that is complementary to either: (1) a sequence of a genomic region; or (2) a sequence that varies from the sequence of (1) only by one or more transitions.

[0264] In some embodiments, MRD is measured via immune repertoire sequencing. TCRs and immunoglobulins serve as a unique “barcode” for these lymphocytes. By sequencing the immune receptors, the immune response in patient samples and clonal expansion of specific lymphocyte subpopulations can be tracked and monitored.

[0265] In some embodiments, MRD is measured by amplifying and sequencing cell-free nucleic acids that are released into bodily fluid from cancer cells, e.g., circulating tumor DNA (ctDNA). In some embodiments, ctDNA can be non-encapsulated tumor-derived fragmented DNA and / or may have one or more epigenetic modifications, for example, the ctDNA may be acetylated, 5-methylated, ubiquitylated, phosphorylated, sumoylated, ribosylated, and / or citrullinated.

[0266] In some embodiments, subjects of the present disclosure undergo tumor biopsies or liquid biopsies and the samples are evaluated for CD22 expression. In some embodiments, subjects of the present disclosure undergo tumor biopsies or liquid biopsies before administration of hypoimmune allogenic CD22- directed CAR T cells of the present disclosure and biopsies are assessed to determine mechanisms of cancer resistance. In some embodiments, subjects of the present disclosure undergo tumor biopsies or liquid biopsies after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure and tumor cells are assessed to determine mechanisms of tumor resistance. Page 64 of 173 12403779v1Attorney Docket No. 2017428-0684

[0267] In some embodiments, PBMCs collected from subjects of the present disclosure before and after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure are assessed for immunophenotypic changes induced by the hypoimmune allogenic CD22-directed CAR T cells of the present disclosure, including expression level of CD47, broad immune cell phenotyping (e.g., lymphoid and myeloid cell phenotypes, e.g., T, B, NK and monocytes), and immune cell subsets (e.g., differentiation, activation, and exhaustion).

[0268] In some embodiments, subjects of the present disclosure are assessed for their survival after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure.

[0269] In some embodiments, subjects of the present disclosure are assessed for certain safety and efficacy criteria after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments the certain safety criteria comprise CRS Grade 3 / 4 ≤10%, ICANS Grade 3 / 4 ≤15%, infections Grade 3 / 4< 15%, and other Grade 3 / 4 adverse events < 5%. In some embodiments the certain safety criteria comprise hypoimmune allogenic CD22-directed CAR T cells of the present disclosure being no worse than autologous CAR T cells. In some embodiments, certain efficacy criteria comprise an event-free survival (EFS) of 10 months, complete response rate (CRR) of 65%, and a progression free survival (PFS) of 15 months for subjects with large B cell lymphoma (LBCL) administered hypoimmune allogenic CD22-directed CAR T cells of the present disclosure as a second-line treatment. In some embodiments, a complete response (CR) is defined as no detectable evidence of tumor. In some embodiments, certain efficacy criteria comprise an overall response rate (ORR) of 70%, CRR of 50%, durable CRR of ≥ 40%, and median duration of response (mDOR) ≥ 12 months for subjects with LBCL administered hypoimmune allogenic CD22-directed CAR T cells of the present disclosure as a third-line treatment. In some embodiments, certain efficacy criteria comprise an ORR of 70%, CRR of 50%, durable CRR of ≥ 40%, and mDOR ≥ 12 months for subjects with transplant ineligible LBCL administered hypoimmune allogenic CD22-directed CAR T cells of the present disclosure as a second-line treatment. In some embodiments, certain efficacy criteria comprise an ORR of 80%, CRR of 40%, durable CRR of 30-40%, mDOR ≥ 12 months, and 70% MRD negativity for subjects with CLL administered hypoimmune allogenic CD22-directed CAR T cells of the present disclosure as a third-line treatment. In some embodiments, certain efficacy criteria comprise ≥50% durable CRR and ≥60% best overall CRR in LBCL patients. In some embodiments, certain efficacy criteria comprise ≥30% best overall CRR and ≥3 months durability in CLL patients. In some embodiments, hypoimmune allogenic CD22-directed CAR T cells of the present disclosure persist for at least 3 months and demonstrate HIP-mediated immune evasion in subjects of the present disclosure. 7. Procedures for Subjects

[0270] In some embodiments, subjects of the present disclosure undergo screening before administration of a lymphodepletion regiment as described herein and / or before administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, screening comprises one or more of: informed consent, demographic data, medical history, previous cancer treatment history, concomitant medications documentation, clinical examination including height and weight, vital signs, neurologic examination (ICE Score), ECOG performance status, ECHO, ECG, brain MRI, lumbar puncture (if clinically indicated), disease assessment (including imaging and bone marrow biopsy / aspirate where applicable), tumor biopsy (if archival tissue not available), SAE reporting, local laboratory assessments (e.g., chemistry panel (including creatinine clearance, as estimated by MDRD equation) (serum), CBC with differential (blood), beta- human chorionic gonadotropin (β-hCG) pregnancy test (serum or urine) for all females of childbearing potential, Page 65 of 173 12403779v1Attorney Docket No. 2017428-0684 serologic tests (serum) (i.e., HIV, hepatitis B virus, and HCV), coagulation (plasma), urinalysis, LDH, CRP, Ferritin,beta-2 microglobulin, direct antiglobulin test, haptoglobin, COVID-19 test, serology (HIV, HBV, HCV), serum immunoglobulin (Ig) levels, immunohistochemistry analysis (biopsy), ctDNA, or MRD.

[0271] In some embodiments, the following lymphodepletion protocol is followed for a subject of the present disclosure. On Days -5 through -3, before the infusion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure, subjects receive a non-myeloablative lymphodepletion (LD) regimen consisting of cyclophosphamide and fludarabine to induce lymphocyte depletion and create an optimal environment for the expansion of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure in vivo. Hypoimmune allogenic CD22-directed CAR T cells of the present disclosure should be administered 3 days (range 2-7 days) after completion of LD chemotherapy.

[0272] In some embodiments, subjects of the present disclosure receive the following 3-day chemotherapy regimen per the order identified as follows: 1) IV hydration with 1 L of 0.9% sodium chloride (NaCl) given before cyclophosphamide on the day of infusion; 2) Cyclophosphamide 500 mg / m2IV over approximately 60 minutes on Days -5, -4, and -3; 3) Fludarabine 30 mg / m2IV over approximately 30 minutes on Days -5, -4, and -3 and subjects with moderate impairment of renal function (creatinine clearance 50-70 mL / min / 1.73 m2) should have their fludarabine dose reduced by 20% and be monitored closely; 4) an additional 1 L of 0.9% NaCl at the completion of the fludarabine infusion; and 5) add Mesna (sodium 2- mercaptoethanesulfonate; a detoxifying agent used to inhibit the hemorrhagic cystitis induced by the chemotherapy) per institutional guidelines. Subjects at risk for tumor lysis syndrome (TLS) should receive TLS prophylaxis prior to LD chemotherapy according to institutional guidelines.

[0273] Administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure can increase the risk of high-grade and fatal toxicities in subjects with ongoing infection and / or inflammation. In some embodiments, if a subject of the present disclosure has: a temperature > 38⁰C within 72 hours before LD chemotherapy; CRP ≥ 100 mg / L any time between enrollment and the start of LD chemotherapy; and / or a white blood cell (WBC) count or WBC differential concerning for infectious process between enrollment and the start of LD chemotherapy (e.g., WBC > 20,000 cells / μL, rapidly increasing WBC, or WBC differential with a high percentage of segments / bands), the cause of the infection should be identified and treated before administration of an LD regimen of the present disclosure.

[0274] In some embodiments, if any screening assessments or procedures are repeated between confirmation of eligibility and the start of LD chemotherapy and the results are outside of the eligibility criteria provided herein, any suspected / identified infectious / inflammatory process condition must resolve before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, a complete medical history and physical examination of a subject of the present disclosure including the head, eyes, ears, nose, and throat (HEENT) and cardiac, vascular, respiratory, gastrointestinal, integumentary, and neurological systems does not reveal evidence of infection / inflammation before administration of an LD regimen of the present disclosure. In some embodiments, a subject of the present disclosure has not received systemic antimicrobials for the treatment of a known or suspected infection within the 48 hours before an LD regimen of the present disclosure (prophylactic use of antimicrobials is allowed). In some embodiments, a treatment course of any antimicrobials given for a known or suspected antecedent infection should be completed before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, if a subject of the present disclosure is confirmed to have an Page 66 of 173 12403779v1Attorney Docket No. 2017428-0684 infectious process for which antimicrobials are not available (e.g., viral pneumonia), the infection must be clinically resolved before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, the most recently collected blood, urine, or other body fluid cultures must show no growth for at least 48 hours, and any other infectious workup performed (e.g., bacterial, PCR, or stool or imaging studies) is negative before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, if there is clinical suspicion of an infection for which cultures are unlikely to be positive within 48 hours (e.g., fungal infection), then adequate time is allowed to allow the cultures to become positive before a subject of the present disclosure is administered an LD regimen of the present disclosure.

[0275] In some embodiments, subjects of the present disclosure are hospitalized during a treatment period from Day 0 before infusion with hypoimmune allogenic CD22-directed CAR T cells of the present disclosure until a minimum of 7 days after infusion. In some embodiments, the volume of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure infused, thaw start / stop time, and the infusion start / stop time must all be noted in a subject’s medical record. In some embodiments, hypoimmune allogenic CD22- directed CAR T cells of the present disclosure are not be thawed until the subject is ready for the infusion.

[0276] In some embodiments, subjects of the present disclosure remain hospitalized until all CAR-T related non-hematologic toxicities resolve to Grade 1 or return to baseline. In some embodiments, subjects may be discharged for a hospital with noncritical and clinically stable or improving toxicities (e.g., renal insufficiency) even if higher than Grade 1. In some embodiments, subjects should remain hospitalized for ongoing hypoimmune allogenic CD22-directed CAR T cells of the present disclosure-related fever, hypotension, hypoxia, or ongoing neurotoxicity of higher than Grade 1. In some embodiments, subjects of the present disclosure refrain from driving or hazardous activities for at least 8 weeks following infusion.

[0277] In some embodiments, after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure and being discharged from the hospital, a subject of the present disclosure will be evaluated at the following intervals: Day 8 (± 1 day), Day 10 (± 1 day), Day 13 (± 1 day), Day 16 (± 1 day), Day 21 (± 4 days), Day 28 (± 4 days), monthly in months 2-12 (± 1 week) (or until start of a new systemic therapy). In some embodiments, subjects undergo the following assessments and procedures at the aforementioned intervals: clinical examination, vital signs, weight, ECOG assessment, concomitant medications documentation, neurologic examination to assess ICE score, ECG (if clinically indicated), brain MRI (if clinically indicated), lumbar puncture (if clinically indicated), tumor biopsy, disease / overall response assessments (including imaging and bone marrow biopsy / aspirate when applicable), AE / SAE / DLT reporting, chemistry panel (serum), CBC with differential (blood), coagulation (plasma), urinalysis (if clinically indicated), LDH, CRP, ferritin, beta-2 microglobulin, direct antiglobulin test, haptoglobin, serum Ig levels, Covid-19 and other viral serology testing (as clinically indicated), β-hCG pregnancy test (serum or urine) for all females of childbearing potential (as clinically indicated), analytes including cytokines (serum / plasma) and anti-CAR-T antibodies, i.e., donor specific antibody and CD22-directed CAR antibody (serum), ctDNA, MRD, CD22-directed CAR T cell kinetics (VCN and FACS), CD47 expression, RCL, T cell clonality, and exploratory analyses including immunophenotyping, T cell killing, NK cell killing, and scRNAseq (PBMCs), assessment of levels of analytes including cytokines (serum / plasma) and CD22-directed CAR T cells (PBMCs) should be drawn at the first onset and first reoccurrence of any Grade 2 or higher CAR-T-related toxicity, such as a Grade 2 CRS or neurologic event, and upon resolution of the event, if not already collected on the day (serum / plasma). Page 67 of 173 12403779v1Attorney Docket No. 2017428-0684

[0278] In some embodiments, a subject of the present disclosure does not receive subsequent anti- cancer therapy. In some embodiments, a subject of the present disclosure does receive subsequent anti-cancer therapy if their disease progression has been documented. In some embodiments, subsequent anti-cancer therapy is or comprises chemotherapy, immunotherapy, targeted agents, SCT, radiation therapy, corticosteroids (other than those used to manage CRS and ICANS), and other investigational agents. 8. Adverse Events and Toxicity Management

[0279] In some embodiments, a subject of the present disclosure experiences one or more adverse events (AEs) after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, an AE is or comprises any untoward medical occurrence in a subject of the present disclosure. In some embodiments, an AE is or comprises worsening of a pre-existing medical condition. Worsening indicates that the pre-existing medical condition has increased in severity, frequency, and / or duration or has an association with a worse outcome. In some embodiments, an AE does not include the following: a pre-existing condition that has not or involves an intervention such as elective cosmetic surgery or a medical procedure; interventions (and associated complications) for pre-treatment conditions (such as elective cosmetic surgery) or medical procedures that were planned before administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure; hospitalization for infusions of hypoimmune allogenic CD22-directed CAR T cells or related procedures, and hospitalization as a precautionary measure. In some embodiments, “disease progression” as assessed by measurement of malignant lesions on radiographs or other methods is not considered to be an AE. In some embodiments, worsening of signs and symptoms of a B cell malignancy of the present disclosure are considered to be AEs.

[0280] In some embodiments, a subject of the present disclosure experiences one or more dose limiting toxicities (DLTs) after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, a DLT is or comprises one or more of the following events that are at least possibly related to treatment with hypoimmune allogenic CD22-directed CAR T cells of the present disclosure and that occur within 28 days of infusion with hypoimmune allogenic CD22-directed CAR T cells of the present disclosure: any Grade 5 event (death) without clinical or radiologic evidence of disease progression; CRS: all Grade 4 CRS, and Grade 3 CRS that fails to improve to ≤ Grade 2 within 72 hours following adequate therapy; Neurotoxicity: All ≥ Grade 3 neurotoxicity of any duration; Grade 3 or higher acute GvHD that requires oral or IV corticosteroids (> 1 mg / kg) and does not resolve within 7 days. Grade 2 acute GvHD that is steroid refractory (i.e., progression after 3 days or no response after 7 days of systemic steroid treatment [≥ 1mg / kg / day prednisone or equivalent]); Grade 4 neutropenia or thrombocytopenia, not attributable to underlying disease or lymphodepletion chemotherapy, that does not improve to ≤ Grade 2 within 42 days. Grade 3 thrombocytopenia with clinically significant bleeding; ≥ Grade 3 toxicity involving vital organs (e.g., cardiac, pulmonary) of any duration (Exceptions may be made for Grade 3 or 4 abnormal hepatic or renal function tests that improves to ≤ Grade 2 within 7 days); all other Grade 3 toxicities, not attributable to underlying disease or lymphodepleting chemotherapy, that do not resolve to ≤ Grade 2 within 72 hours. In some embodiments, the following events are not considered to be DLTs: Grade 3 cytopenia or Grade 4 lymphopenia / anemia, Grade 3 TLS for ≤ 2 weeks, and / or Grade 3 infections.

[0281] In some embodiments, a subject of the present disclosure experiences one or more serious adverse events (SAEs) after administration of hypoimmune allogenic CD22-directed CAR T cells of the present disclosure. In some embodiments, an SAE is defined as an AE that meets at least one of the following serious Page 68 of 173 12403779v1Attorney Docket No. 2017428-0684 criteria: is fatal, is life-threatening (i.e., an event that places the subject at immediate risk of death; it does not refer to an event that hypothetically might have caused death if it were more severe), requires inpatient hospitalization or prolongation of existing or planned hospitalization, an AE would meet the criterion of “requires hospitalization” if the event necessitated an admission to a healthcare facility, events that require an escalation of care when the subject is already hospitalized should be recorded as an SAE (e.g., movement from routine care in the hospital to the intensive care unit or if that event resulted in a prolongation of the planned hospitalization, results in persistent or significant disability / incapacity, is a congenital anomaly / birth defect, and / or is another medically important serious event. The terms “severe” and “serious” are not synonymous. Severity refers to the intensity of an AE according to NCI Common Terminology Criteria for Adverse Events (CTCAE); the event itself may be of relatively minor medical significance and, therefore, may not meet the seriousness criteria listed above. Severity and seriousness need to be independently assessed for each AE. Progression of a malignancy is not considered to be a drug-related SAE; signs and symptoms of disease progression can be considered to be SAEs (and documented as being due to disease progression).

[0282] In some embodiments, an AE is or comprises a targeted AE. In some embodiments, an SAE is or comprises a targeted SAE. Targeted AEs / SAEs include neurologic, hematologic, infection, GvHD, autoimmune disorder, and second primary malignancy events. Adverse events of special interest (reflecting potential risks of CD22-directed CAR therapies) include: CRS, neurotoxicity (i.e., ICANS), Macrophage Activation Syndrome, TLS, Grade 3 cytopenia persisting through Day 28 post-treatment, Severe (Grade 3) infection through the first 28 days post-treatment.

[0283] In some embodiments, AEs are determined based on abnormal clinical laboratory findings. In some embodiments, abnormal laboratory findings without clinical significance are not AEs. In some embodiments, abnormal laboratory findings that result in new or worsening clinical sequelae or that require therapy or adjustment in current therapy are considered AEs. In some embodiments, an AE is an abnormal laboratory test result that results in a medical intervention (e.g., potassium supplementation for hypokalemia or iron replacement therapy for anemia) or a change in concomitant therapy. In some embodiments, an abnormal vital sign results is an AE if it is a change from baseline and is accompanied by clinical symptoms and / or results in a medical intervention or a change in concomitant therapy.

[0284] In some embodiments, risks associated with CAR T cell treatment are or comprise cytokine release syndrome (CRS), neurotoxicity (i.e., ICANS), cytopenia, hypogammaglobulinemia and infections, T cell lymphoma, and TLS. In some embodiments, risks associated with allogeneic CAR T cell therapy include GvHD and clonal expansion (e.g., second primary malignancy).

[0285] In some embodiments, if a subject of the disclosure experiences CRS, it is managed according to ASTCT guidelines (Lee 2019). In some embodiments, if a subject of the disclosure experiences neurotoxicity (i.e., ICANS) it is managed according to ASTCT guidelines (Lee 2019). In some embodiments, if a subject of the disclosure experiences GvHD it is managed according to Mount Sinai Acute GVHD International Consortium 2016 report (Harris 2016).

[0286] Infections, hypogammaglobulinemia, and cytopenia are known side effects of CAR T cell therapy. Infections after CAR T cell therapy are common and have been reported in up to 70% of patients who received CAR T cell therapy in registrational clinical trials for approved agents (KYMRIAH® Prescribing Information, YESCARTA® Prescribing Information, BREYANZI® Prescribing Information). Bacterial, viral, and Page 69 of 173 12403779v1Attorney Docket No. 2017428-0684 fungal infections have all been reported with use of CAR T cell therapy. Infections may occur for a number of concomitant reasons, including lymphodepleting (or antecedent) chemotherapy, CAR T cell–mediated B cell or plasma cell depletion, prolonged cytopenia, corticosteroid treatment, or as a consequence of the malignancy itself. The severity of CRS may also be associated with an increased risk of acute infections. Other potential risk factors for severe infections within the first 30 days of CAR-T treatment include ICANS, tocilizumab, and corticosteroid use. Subjects may remain at increased risk of complications for weeks to months after infusion (Logue 2021, Hill 2018). Infections are generally managed using agents that target the source of infection. Additionally, prophylaxis against vesicular stomatitis virus / herpes simplex virus reactivation and neumocystis jirovecii pneumonia infections is generally used for patients undergoing CAR T cell therapy and for several months after. The decision to administer antibacterial or antifungal prophylaxis should be risk-adjusted based on subject characteristics, such as prior lines of suppressive therapy and infection history (Thompson 2022).

[0287] Hypogammaglobulinemia is characterized by low antibody levels due to extremely low B cell or plasma cell counts, referred to as B cell or plasma cell aplasia, respectively. Hypogammaglobulinemia has been reported in up to 53% of patients who received CAR T cell therapy in registrational clinical trials and is associated with risk of infection (KYMRIAH® Prescribing Information, YESCARTA® Prescribing Information, BREYANZI® Prescribing Information). After lymphodepleting chemotherapy and treatment of the present disclosure, subjects can be monitored for serum immunoglobulin (Ig) levels and CBC with differential. Subjects with hypogammaglobulinemia (serum IgG levels < 400–600 mg / dL) and serious or recurrent infections (particularly bacterial) should be managed with intravenous immunoglobulin (IVIG) replacement therapy (400-500 mg / kg IVIG given monthly). IVIG should be continued until serum IgG levels normalize and infections are resolved. The optimal IgG threshold to use may depend on subject characteristics and infection frequency or severity.

[0288] Subjects who receive CAR T cell therapy are also at risk for hematologic toxicities, including prolonged cytopenia, such as neutropenia, thrombocytopenia, anemia, and / or leukopenia (KYMRIAH® Prescribing Information, YESCARTA® Prescribing Information, BREYANZI® Prescribing Information). Cytopenia may occur in the weeks to months after lymphodepleting chemotherapy and CAR T cell therapy infusion. Factors that may contribute to prolonged cytopenia include CRS and ICANS severity, disease burden, the number of prior therapies, baseline blood cell counts, peak CRP and ferritin levels, and CAR construct. Although lymphodepletion may be a contributing factor, the pathophysiology of prolonged cytopenia after CAR T cell infusion remains unclear. Alternative causes of cytopenia in treated include myelodysplastic syndrome or second primary malignancy (Jain 2020, Fried 2019). Cytopenia are generally managed with transfusion or growth factor support, if the possibility of myelodysplastic syndrome has been ruled out. Growth factors may be considered for persistent cytopenia. GM-CSF is not recommended in the setting of CAR T cell therapy. An FDA-approved biosimilar is an appropriate substitute for filgrastim (Thompson 2022). B. Hypoimmunogenic Cells

[0289] Allogeneic immune recognition and rejection remain an obstacle in cellular transplantation and cell therapy development. Abrogation of HLA class I and II function and overexpression of CD47 are important genetic modifications to evade immune recognition. CD47, a membrane protein that interacts with signal regulatory protein-alpha (SIRPα) on innate immune cells, is a key protein for evading immune recognition by innate immune cells, including natural killer cells (NK cells) and macrophages, which can immediately kill Page 70 of 173 12403779v1Attorney Docket No. 2017428-0684 allogeneic cells. Hematologic cells, including red blood cells, upregulate CD47 to avoid innate immune cell recognition and destruction.

[0290] Allogeneic CAR T cells of the present disclosure involve impairment of HLA class I / II function through knockout of beta-2 microglobulin (B2M) and class II major histocompatibility complex transactivator (CIITA). While such HLA impairment can result in adaptive immune evasion, innate reactivity can be enhanced. Therefore, CAR T cells of the present disclosure additionally overexpress CD47 in order to evade this potential innate immune response.

[0291] During development, NK cells undergo an educational “licensing” process to ensure that only those NK cells that express a cognate inhibitory receptor for self-HLA class I molecules become functionally mature. This central self-tolerance mechanism sets a triggering threshold, and the integration of all transmitted activating and inhibitory signals can determine the outcome and magnitude of interactions with target cells. In- depth analysis of NK cell interactions with immune-edited cells indicates that SIRPα serves as an immune checkpoint molecule by delivering a strong inhibitory signal when engaging with its cognate ligand, CD47. An immune engineering strategy including CD47 may thus provide protection against macrophages but also NK cell killing. Without wishing to be bound by any particular theory, data suggests that that these genetic modifications do not induce immunological tolerance but confer a lack of immune recognition of the transplanted cells.

[0292] In some embodiments, the present disclosure is directed primary cells (such as, but not limited to, primary T cells). In some embodiments, primary cells such as primary T cells, are engineered for reduced expression or lack of expression of human leukocyte antigen (HLA) class I and / or HLA class II human leukocyte antigens, and in some instances, for reduced expression or lack of expression of a T-cell receptor (TCR) complex. In some embodiments, the hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a chimeric antigen receptor (CAR) in addition to reduced expression or lack of expression of HLA class I and / or HLA class II human leukocyte antigens, and have reduced expression or lack expression of a T-cell receptor (TCR) complex. In some embodiments, the CAR is a CD22-directed CAR. In some embodiments, the cells are modified or engineered as compared to a wild-type or control cell, including an unaltered or unmodified wild-type cell or control cell. In some embodiments, the wild-type cell or the control cell is a starting material. In some embodiments, the starting material is a primary cell collected from a donor. In some embodiments, the starting material is a primary blood cell collected from a donor, e.g., via a leukopak. In some embodiments, the starting material is otherwise modified or engineered to have altered expression of one or more genes to generate the engineered cell.

[0293] In some embodiments, engineered and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a chimeric antigen receptor (CAR), and include a genomic modification of the B2M gene. In some embodiments, engineered and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and include a genomic modification of the CIITA gene. In some embodiments, engineered and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a CAR, and include a genomic modification of the TRAC gene. In some embodiments, engineered and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a CAR, and include one or more genomic modifications selected from the group consisting of the B2M, CIITA, and TRAC genes. In some embodiments, engineered and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a CAR, and include genomic modifications of the B2M, CIITA, and TRAC genes. In some embodiments, the cells are B2M- / -, CIITA- / -, TRAC- / -, CD47tg cells that also express CARs. Page 71 of 173 12403779v1Attorney Docket No. 2017428-0684

[0294] In certain embodiments, the cells are B2Mindel / indel, CIITAindel / indel, TRACindel / indel, CD47tg cells that also express CARs. Non-limiting examples of primary T cells include CD3+ T cells, CD4+ T cells, CD8+ T cells, naïve T cells, regulatory T (Treg) cells, non-regulatory T cells, Th1 cells, Th2 cells, Th9 cells, Th17 cells, T-follicular helper (Tfh) cells, cytotoxic T lymphocytes (CTL), effector T (Teff) cells, central memory T (Tcm) cells, effector memory T (Tem) cells, effector memory T cells express CD45RA (TEMRA cells), tissue- resident memory (Trm) cells, virtual memory T cells, innate memory T cells, memory stem cell (Tsc), γδ T cells, and any other subtype of T cells. In some embodiments, primary T cells are selected from a group that includes cytotoxic T-cells, helper T-cells, memory T-cells, regulatory T-cells, tumor infiltrating lymphocytes, and combinations thereof. In some embodiments, cells are modified or engineered as compared to a wild-type or control cell, including an unaltered or unmodified wild-type cell or control cell. In some embodiments, a wild-type cell or a control cell is a starting material. In some embodiments, starting material is a primary cell collected from a donor. In some embodiments, the donor is a healthy donor. In some embodiments, starting material is a primary blood cell collected from a donor, e.g., via a leukopak. In some embodiments, starting material is otherwise modified or engineered to have altered expression of one or more genes to generate an engineered cell.

[0295] In some embodiments, primary T cells are from a pool of primary T cells from one or more donor subjects that are different than the recipient subject (e.g., the patient administered the cells). Primary T cells can be obtained from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100 or more donor subjects and pooled together. Primary T cells can be obtained from 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10, or more 20 or more, 50 or more, or 100 or more donor subjects and pooled together. In some embodiments, primary T cells are harvested from one or a plurality of individuals, and in some instances, primary T cells or a pool of primary T cells are cultured in vitro. In some embodiments, primary T cells or a pool of primary T cells are engineered to exogenously express CD47 and cultured in vitro.

[0296] In certain embodiments, primary T cells or a pool of primary T cells are engineered to express a chimeric antigen receptor (CAR). Useful CARs include those that bind a CD22 antigen.

[0297] In some embodiments, the primary T cells or the pool of primary T cells are engineered to exhibit reduced expression of an endogenous T cell receptor compared to unmodified primary T cells. Methods of genetically modifying a cell including a T cell are described in detail, for example, in WO2020 / 018620 and WO2016 / 183041, the disclosures of which are herein incorporated by reference in their entireties, including the tables, appendices, sequence listing and figures.

[0298] In some embodiments, CAR-T cells comprise a second generation CAR comprising an antigen binding domain, a transmembrane domain, and at least two signaling domains.

[0299] In some embodiments, CAR-T cells comprise a CAR comprising an antigen binding domain, a transmembrane, and one or more signaling domains. In some embodiments, the CAR also comprises a linker. In some embodiments, the CAR comprises a CD22 antigen binding domain. In some embodiments, the CAR comprises a CD8α transmembrane domain. In some embodiments, the CAR comprises a CD8α signal peptide. In some embodiments, the CAR comprises a Whitlow linker GSTSGSGKPGSGEGSTKG (SEQ ID NO: 38). In some embodiments, the antigen binding domain of the CAR is selected from a group including, but not limited to, (a) an antigen binding domain targets an antigen characteristic of a neoplastic cell or an antigen binding domain targets an antigen characteristic of an autoimmune disorder. Page 72 of 173 12403779v1Attorney Docket No. 2017428-0684

[0300] In some embodiments, the CAR further comprises one or more linkers. The format of an scFv is generally two variable domains linked by a flexible peptide sequence, or a “linker,” either in the orientation VH- linker-VL or VL-linker-VH. Any suitable linker known to those in the art in view of the specification can be used in the CARs. Examples of suitable linkers include, but are not limited to, a GS based linker sequence, a G4S linker sequence (SEQ ID NO: 37), and a Whitlow linker GSTSGSGKPGSGEGSTKG (SEQ ID NO: 38).

[0301] In some embodiments, the antigen binding domain is selected from a group that includes an antibody, an antigen-binding portion or fragment thereof, an scFv, and a Fab. In some embodiments, the antigen binding domain binds to CD22. In some embodiments, the antigen binding domain is an anti-CD22 scFv such as but not limited to m971.

[0302] In some embodiments, the transmembrane domain comprises one selected from a group that includes a transmembrane region of TCRα, TCRβ, TCRζ, CD3ε, CD3γ, CD3δ, CD3ζ, CD4, CD5, CD8α, CD8β, CD9, 4-. a can a a domain can contain one or more costimulatory domains. In certain embodiments, the signaling domain comprises a costimulatory domain. In other embodiments, the signaling domains comprise costimulatory domains. In some cases, when the CAR comprises two or more costimulatory domains, two costimulatory domains are not the same. In some embodiments, the costimulatory domains comprise two costimulatory domains that are not the same. In some embodiments, the costimulatory domain enhances cytokine production, CAR-T cell proliferation, and / or CAR-T cell persistence during T cell activation. In some embodiments, the costimulatory domains enhance cytokine production, CAR-T cell proliferation, and / or CAR-T cell persistence during T cell activation.

[0304] In some embodiments, a CAR comprises a CD3 zeta (CD3ζ) intracellular domain or functionalskilled in the art. Detailed descriptions are found, for example, in Vormittag et al., Curr Opin Biotechnol, 2018, 53, 162-181; and Eyquem et al., Nature, 2017, 543, 113-117.

[0306] In some embodiments, the cells derived from primary T cells comprise reduced expression of an endogenous T cell receptor, for example by disruption of an endogenous T cell receptor gene (e.g., T cell receptor alpha constant region (TRAC)).

[0307] In some embodiments, a CD47 transgene is inserted into a random locus of a cell. In some embodiments, a transgene encoding a CAR is inserted into a random locus of a cell. In some embodiments, a transgene encoding a CAR is inserted into a random locus of a cell via viral vector transduction / integration. In some embodiments, a CD47 transgene and a transgene encoding a CAR are inserted into a random locus of a cell via viral vector transduction / integration. In some embodiments, a vector is a self-inactivating lentiviral vector pseudotyped with a vesicular stomatitis VSV-G envelope. In some embodiments, a transgene encoding a CAR is Page 73 of 173 12403779v1Attorney Docket No. 2017428-0684 inserted into at least one allele of a cell using viral transduction. In some embodiments, an exogenous polynucleotide is inserted into at least one allele of a cell using a lentivirus based viral vector.

[0308] In some embodiments, a CD47 transgene and a transgene encoding a CAR are inserted into the same locus.

[0309] In certain embodiments, a CD47 and a CAR are controlled by a single promoter and are encoded by a single transgene. In some instances, the promoter controlling expression of any transgene described is a constitutive promoter. In other instances, the promoter for any transgene described is an inducible promoter. In some embodiments, the promoter is an EF1α promoter. In some embodiments, the promoter is CAG promoter. In some embodiments, a CD47 transgene and a transgene encoding a CAR are both controlled by a constitutive promoter. In some embodiments, a CD47 transgene and a transgene encoding a CAR are both controlled by an inducible promoter.

[0310] Methods provided are useful for inactivation or ablation of HLA class I expression and / or HLA class II expression in cells such as primary T cells. In some embodiments, genome editing technologies utilizing rare-cutting endonucleases (e.g., CRISPR / Cas) are also used to reduce or eliminate expression of genes involved in an immune response (e.g., by deleting genomic DNA of genes involved in an immune response or by insertions of genomic DNA into such genes, such that gene expression is impacted) in cells. In certain embodiments, genome editing technologies or other gene modulation technologies are used to insert tolerance- inducing factors in human cells, rendering them and the differentiated cells prepared therefrom hypoimmunogenic cells. As such, the hypoimmunogenic cells have reduced or eliminated expression of HLA I and HLA II expression. In some embodiments, the cells are nonimmunogenic (e.g., do not induce an innate and / or an adaptive immune response) in a recipient subject.

[0311] In some embodiments, a cell includes a modification to increase expression of CD47.

[0312] In some embodiments, a cell comprises a genomic modification of one or more target polynucleotide sequences that regulate the expression of either HLA class I molecules, HLA class II molecules, or HLA class I and HLA class II molecules. In some embodiments, a genetic editing system is used to modify one or more target polynucleotide sequences. In some embodiments, the targeted polynucleotide sequence is one or more selected from the group including B2M and CIITA. In some embodiments, the cell comprises a genetic editing modification to the B2M gene. In some embodiments, the cell comprises a genetic editing modification to the CIITA gene. In some embodiments, the cell comprises genetic editing modifications to the B2M and CIITA genes. In certain embodiments, the genome of the cell has been altered to reduce or delete critical components of HLA expression. In some embodiments, the cells are modified or engineered as compared to a wild-type or control cell, including an unaltered or unmodified wild-type cell or control cell. In some embodiments, the wild- type cell or the control cell is a starting material. In some embodiments, the starting material is a primary cell collected from a donor. In some embodiments, the starting material is a primary blood cell collected from a donor, e.g., via a leukopak. In some embodiments, the starting material is otherwise modified or engineered to have altered expression of one or more genes to generate the engineered cell.

[0313] In some embodiments, the present disclosure provides a cell (e.g., primary T cell or CAR-T cell) or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of HLA class I molecules in the cell or population thereof. In certain embodiments, the present disclosure provides a cell (e.g., primary T cell or CAR-T Page 74 of 173 12403779v1Attorney Docket No. 2017428-0684 cell) or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of HLA class II molecules in the cell or population thereof. In numerous embodiments, the present disclosure provides a cell (e.g., primary T cell or CAR-T cell) or population thereof comprising a genome in which one or more genes has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of HLA class I and II molecules in the cell or population thereof.

[0314] In certain embodiments, the expression of HLA I molecules and / or HLA II molecules is modulated by targeting and deleting a contiguous stretch of genomic DNA, thereby reducing or eliminating expression of a target gene selected from the group consisting of B2M and CIITA. In some embodiments, described herein are genetically edited cells (e.g., modified human cells) comprising exogenous CD47 proteins and inactivated or modified CIITA gene sequences, and in some instances, additional gene modifications that inactivate or modify B2M gene sequences.

[0315] Provided herein are cells exhibiting a modification of one or more targeted polynucleotide sequences that regulates the expression of any one of the following: (a) HLA I antigens, (b) HLA II antigens, (c) TCR complexes, (d) both HLA I and II antigens, and (e) HLA I and II antigens and TCR complexes. In certain embodiments, the modification includes increasing expression of CD47. In some embodiments, the cells include an exogenous or recombinant CD47 polypeptide. In certain embodiments, the modification includes expression of a chimeric antigen receptor. In some embodiments, the cells comprise an exogenous or recombinant chimeric antigen receptor polypeptide.

[0316] In some embodiments, the present disclosure provides a cell or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of HLA class I molecules in the cell or population thereof. In certain embodiments, the present disclosure provides a cell or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of HLA class II molecules in the cell or population thereof. In certain embodiments, the present disclosure provides a cell or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of TCR molecules in the cell or population thereof. In numerous embodiments, the present disclosure provides a cell or population thereof comprising a genome in which one or more genes has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of HLA class I and II molecules and TCR complex molecules in the cell or population thereof.

[0317] In some embodiments, the cells and methods described herein include genomically editing human cells to cleave CIITA gene sequences as well as editing the genome of such cells to alter one or more additional target polynucleotide sequences such as, but not limited to, B2M and TRAC. In some embodiments, the cells and methods described herein include genomically editing human cells to cleave B2M gene sequences as well as editing the genome of such cells to alter one or more additional target polynucleotide sequences such as, but not limited to, CIITA and TRAC. In some embodiments, the cells and methods described herein include genomically editing human cells to cleave TRAC gene sequences as well as editing the genome of such cells to alter one or more additional target polynucleotide sequences such as, but not limited to, B2M and CIITA. Page 75 of 173 12403779v1Attorney Docket No. 2017428-0684

[0318] In some embodiments, a population of engineered cells described evades NK cell mediated cytotoxicity upon administration to a recipient patient. In some embodiments, a population of engineered cells evades NK cell mediated cytotoxicity by one or more subpopulations of NK cells. In some embodiments, a population of engineered is protected from cell lysis by NK cells, including immature and / or mature NK cells upon administration to a recipient patient. In some embodiments, a population of engineered cells evades macrophage engulfment upon administration to a recipient patient. In some embodiments, a population of engineered cells does not induce an innate and / or an adaptive immune response to the cell upon administration to a recipient patient.

[0319] In some embodiments, cells described herein are controlled by a safety switch. The term “safety switch” used herein refers to a system for controlling the expression of a gene or protein of interest that, when downregulated or upregulated, leads to clearance or death of the cell, e.g., through recognition by the host’s immune system. A safety switch can be designed to be triggered by an exogenous molecule in case of an adverse clinical event. A safety switch can be engineered by regulating the expression on the DNA, RNA and protein levels. A safety switch includes a protein or molecule that allows for the control of cellular activity in response to an adverse event. In some embodiments, a safety switch comprises a therapeutic agent that inhibits or blocks the interaction of CD47 and SIRPα. In some embodiments, a CD47-SIRPα blockade agent is an agent that neutralizes, blocks, antagonizes, or interferes with the cell surface expression of CD47, SIRPα, or both. In some embodiments, the CD47-SIRPα blockade agent inhibits or blocks the interaction of CD47, SIRPα or both. In some embodiments, a CD47-SIRPα blockade agent (e.g., a CD47-SIRPα blocking, inhibiting, reducing, antagonizing, neutralizing, or interfering agent) comprises an agent selected from a group that includes an antibody or fragment thereof that binds CD47, a bispecific antibody that binds CD47, an immunocytokine fusion protein that bind CD47, a CD47 containing fusion protein, an antibody or fragment thereof that binds SIRPα, a bispecific antibody that binds SIRPα, an immunocytokine fusion protein that bind SIRPα, an SIRPα containing fusion protein, and a combination thereof.

[0320] In some embodiments, the population of engineered cells described elicits a reduced level of immune activation or no immune activation upon administration to a recipient subject. In some embodiments, the cells elicit a reduced level of systemic TH1 activation or no systemic TH1 activation in a recipient subject. In some embodiments, the cells elicit a reduced level of immune activation of peripheral blood mononuclear cells (PBMCs) or no immune activation of PBMCs in a recipient subject. In some embodiments, the cells elicit a reduced level of donor-specific IgG antibodies or no donor specific IgG antibodies against the cells upon administration to a recipient subject. In some embodiments, the cells elicit a reduced level of IgM and IgG antibody production or no IgM and IgG antibody production against the cells in a recipient subject. In some embodiments, the cells elicit a reduced level of cytotoxic T cell killing of the cells upon administration to a recipient subject.

[0321] In some embodiments, a hypoimmunogenic cell disclosed herein includes: (i) overexpression of CD47; (ii) B2M and CIITA knock-out; (iii) disruption of TRAC expression; and (iv) expression of a CD22-directed CAR. Without wishing to be bound by any particular theory, CD47 overexpression can serve to block host innate immunity, disruption of HLA Class I / II function via B2M and CIITA knockout can serve to prevent host adaptive immunity, and TRAC disruption can serve to prevent GvHD. 1. CIITA Page 76 of 173 12403779v1Attorney Docket No. 2017428-0684

[0322] In some embodiments, the technologies disclosed herein modulate (e.g., reduces or eliminates) the expression of HLA II genes by targeting and modulating (e.g., reducing or eliminating) Class II transactivator (CIITA) expression. In some embodiments, the modulation occurs using a CRISPR / Cas system. CIITA is a member of the LR or nucleotide binding domain (NBD) leucine-rich repeat (LRR) family of proteins and regulates the transcription of HLA II by associating with the HLA enhanceosome.

[0323] In some embodiments, the target polynucleotide sequence of the present disclosure is a variant of CIITA. In some embodiments, the target polynucleotide sequence is a homolog of CIITA. In some embodiments, the target polynucleotide sequence is an ortholog of CIITA.

[0324] In some embodiments, reduced or eliminated expression of CIITA reduces or eliminates expression of one or more of the following HLA class II are HLA-DP, HLA-DM, HLA-DOA, HLA-DOB, HLA-DQ, and HLA-DR.

[0325] In some embodiments, the cells described herein comprise gene modifications at the gene l...

Claims

Attorney Docket No. 2017428-0684 CLAIMS 1. A method of treating a subject having or suspected of having a B cell malignancy or a B cell mediated disorder, the method comprising: administering to the subject a composition comprising a population of engineered hypoimmunogenic T cells, wherein the hypoimmunogenic T cells comprise one or more modifications that: (a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and / or (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules, (b) increase expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not comprise the modification, and (c) express a CD22-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.

2. A method of treating a subject having or suspected of having a B cell malignancy or a B cell mediated disorder, the method comprising: administering to the subject a composition comprising a population of engineered hypoimmunogenic T cells comprising: (a) reduced expression of beta-2 microglobulin (B2M), Class II Major Histocompatability Complex Transactivator (CIITA), and T cell receptor alpha constant (TRAC) relative to a control or wild-type T cells, (b) increased expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to the control or wild-type T cells, and (c) expression of a CD22-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.

3. The method of claim 1 or claim 2, wherein the composition is administered to the subject at a dosage of about 5x106cells to about 400x106cells.

4. The method of any one of the claims 1-3, wherein the composition is administered to the subject at a dosage of at least about 30x106cells, at least about 60x106cells, at least about 90x106cells, at least about 120x106cells, at least about 150x106cells, at least about 200x106cells, at least about 250x106cells, at least about 300x106cells, at least about 350x106cells, or at least about 400x106cells.

5. The method of any one of claims 1-4, wherein the composition is administered to the subject at a dosage of about 5x106CAR+ cells to about 400x106CAR+ cells. Page 165 of 173 12403779v1Attorney Docket No. 2017428-0684 6. The method of any one of claims 1-5, wherein the composition is administered to the subject at a dosage of at least about 30x106CAR+ cells, at least about 60x106CAR+ cells, at least about 90x106CAR+ cells, at least about 120x106CAR+ cells, at least about 150x106CAR+ cells, at least about 200x106CAR+ cells, at least about 250x106CAR+ cells, at least about 300x106CAR+ cells, at least about 350x106CAR+ cells, or at least about 400x106CAR+ cells.

7. The method of any one of claims 1-6, wherein a total volume of about 1.25mL to about 100mL of the composition is administered.

8. The method of any one of claims 1-7, wherein the composition comprises a concentration of engineered hypoimmunogenic T cells of at least 1.25x106cells / mL.

9. The method of any one of claims 1-8, wherein the CAR comprises: (i) an anti-CD22 antigen binding domain; (ii) a CD8α hinge domain, a CD28 hinge domain, or an IgG4 hinge domain; (iii) a CD8α transmembrane domain, or a CD28 transmembrane domain; (iv) a 4-1BB costimulatory domain, or a CD28 costimulatory domain; and (v) a CD3ζ intracellular signaling domain.

10. The method of claim 9, wherein the antigen binding domain comprises: (i) a light chain amino acid sequence comprising a light chain CDR1, a light chain CDR2, and a light chain CDR3, wherein the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 25, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 26, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 27, and (ii) a heavy chain amino acid sequence comprising a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3 amino acid, wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 21, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 22, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:

23.

11. The method of claim 10, wherein the light chain amino acid sequence comprises a light chain variable region and the heavy chain amino acid sequence comprises a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence at least 90% identical to SEQ ID NO: 24, and wherein the heavy chain variable region comprises an amino acid sequence comprises an amino acid sequence at least 90% identical to SEQ ID NO:

20.

12. The method of any one of claims 9-11, wherein the antigen binding domain comprises a linker comprising an amino acid sequence at least 90% identical to SEQ ID NO:

37. Page 166 of 173 12403779v1Attorney Docket No. 2017428-0684 13. The method of any one of claims 9-12, wherein the antigen binding domain comprises a linker comprising the amino acid sequence of SEQ ID NO:

37.

14. The method of any one of claims 9-13, wherein the antigen binding domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

19.

15. The method of any one of claims 9-14, wherein the antigen binding domain comprises the amino acid sequence of SEQ ID NO:

19.

16. The method of claim 9, wherein the antigen binding domain comprises: (i) a light chain amino acid sequence comprising a light chain CDR1, a light chain CDR2, and a light chain CDR3, wherein the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 34, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 35, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 36, and (ii) a heavy chain amino acid sequence comprising a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3 amino acid, wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 30, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 31, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:

32.

17. The method of claim 16, wherein the light chain amino acid sequence comprises a light chain variable region and the heavy chain amino acid sequence comprises a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence at least 90% identical to SEQ ID NO: 33, and wherein the heavy chain variable region comprises an amino acid sequence comprises an amino acid sequence at least 90% identical to SEQ ID NO:

29.

18. The method of claim 16 or 17, wherein the antigen binding domain comprises a linker comprising an amino acid sequence at least 90% identical to SEQ ID NO:

37.

19. The method of any one of claims 16-18, wherein the antigen binding domain comprises a linker comprising the amino acid sequence of SEQ ID NO:

37.

20. The method of any one of claims 16-19, wherein the antigen binding domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

28.

21. The method of any one of claims 16-20, wherein the antigen binding domain comprises the amino acid sequence of SEQ ID NO:

28. Page 167 of 173 12403779v1Attorney Docket No. 2017428-0684 22. The method of any one of claims 9-21, wherein the CD8α hinge domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

9.

23. The method of any one of claims 9-22, wherein the CD8α hinge domain comprises an amino acid sequence of SEQ ID NO:

9.

24. The method of any one of claims 9-21, wherein the CD28 hinge domain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 10 or SEQ ID NO:

113.

25. The method of any one of claims 9-21 and 24, wherein the CD28 hinge domain comprises an amino acid sequence of SEQ ID NO: 10 or SEQ ID NO:

113.

26. The method of any one of claims 9-21, wherein the IgG4 hinge domain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO:

13.

27. The method of any one of claims 9-21 and 26, wherein the IgG4 hinge domain comprises an amino acid sequence of SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO:

13.

28. The method of any one of claims 9-27, wherein the CD8α transmembrane domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

14.

29. The method of any one of claims 9-28, wherein the CD8α transmembrane domain comprises an amino acid sequence of SEQ ID NO:

14.

30. The method of any one of claims 9-27, wherein the CD28 transmembrane domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

15.

31. The method of any one of claims 9-27 and 30, wherein the CD28 transmembrane domain comprises an amino acid sequence of SEQ ID NO:

15.

32. The method of any one of claims 9-31, wherein the 4-1BB costimulatory domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

16.

33. The method of any one of claims 9-32, wherein the 4-1BB costimulatory domain comprises an amino acid sequence of SEQ ID NO:

16.

34. The method of any one of claims 9-31, wherein the CD28 costimulatory domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

17. Page 168 of 173 12403779v1Attorney Docket No. 2017428-0684 35. The method of any one of claims 9-31 and 34, wherein the CD28 costimulatory domain comprises an amino acid sequence of SEQ ID NO:

17.

36. The method of any one of claims 9-35, wherein the CD3ζ intracellular signaling domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

18.

37. The method of any one of claims 9-36, wherein the CD3ζ intracellular signaling domain comprises an amino acid sequence of SEQ ID NO:

18.

38. The method of any one of claims 1-8, wherein the CAR comprises an amino acid sequence at least 90% identical to SEQ ID NO:

40.

39. The method of claim 38, wherein the CAR comprises an amino acid sequence identical to SEQ ID NO:

40.

40. The method of any one of claims 1-39, wherein the engineered hypoimmunogenic T cells are B2Mindel / indel, CIITAindel / indel, and / or TRACindel / indelcells. . e met od o any one o caims 1-40, wherein the B cell malignancy is a lymphoma or a leukemia.

42. The method of any one of claims 1-41, wherein the B cell malignancy is selected from the group consisting of: Non-Hodgkin’s Lymphoma (NHL), Chronic Lymphocytic Leukemia (CLL), large B cell lymphoma (LBCL), diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), primary mediastinal B cell lymphoma (PMBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), marginal zone lymphoma (MZL), or small lymphocytic lymphoma (SLL).

43. The method of any one of claims 1-42, wherein prior to administration of the composition, CD22+ B Cells are detectable in the circulation of the subject.

44. The method of any one of claims 1-43, wherein prior to administration of the composition, the subject exhibits clinical evidence of B cell malignancy or B cell mediated disorder relapse.

45. The method of claim 44, wherein the clinical evidence comprises lymphadenopathy.

46. The method of any one of claims 1-45, wherein prior to administration of the composition, the subject has cancer cells that do not express CD19.

47. The method of any one of claims 1-46, wherein the B cell malignancy or the B cell mediated disorder is relapsed or refractory following at least one therapeutic standard of care regimen.

48. The method of any one of claims 1-47, wherein the B cell malignancy or the B cell mediated disorder is relapsed or refractory following at least two therapeutic standard of care regimens. Page 169 of 173 12403779v1Attorney Docket No. 2017428-0684 49. The method of any one of claims 1-48, wherein the B cell malignancy or the B cell mediated disorder is relapsed or refractory following at least three therapeutic standard of care regimens.

50. The method of any one of claims 1-49, wherein the B cell malignancy or the B cell mediated disorder is relapsed or refractory following treatment with a CD19-directed CAR cell therapy.

51. The method of any one of claims 1-50, wherein the B cell malignancy or the B cell mediated disorder is relapsed or refractory following autologous stem cell transplant (ASCT).

52. The method of any one of claims 1-51, wherein the B-cell malignancy or B cell mediated disorder is relapsed and / or refractory.

53. The method of claim 52, wherein the B cell malignancy is relapsed and / or refractory NHL.

54. The method of claim 52, wherein the B cell malignancy is relapsed and / or refractory CLL.

55. The method of any one of claims 52-54, wherein the B cell malignancy is relapsed and / or refractory after the subject has received at least one therapeutic regimen.

56. The method of claim 55, wherein the at least one therapeutic regimen comprises a CD19-directed CAR T therapy.

57. The method of claim 55, wherein the at least one therapeutic regimen does not comprise more than one CD19-directed CAR T therapy.

58. The method of claim 56, wherein the CD19-directed CAR T therapy is autologous or allogeneic.

59. The method of any one of claims 56 or 68, wherein the composition is administered at least 3 months after the subject received the CD19-directed CAR T therapy.

60. The method of any one of claims 1-59, wherein the subject is 18-85 years of age.

61. The method of any one of claims 1-60, wherein the method further comprises: administering to the subject a lymphodepleting chemotherapy regimen, wherein the step of administering the lymphodepleting chemotherapy regimen occurs prior to the step of administering the composition.

62. The method of claim 61, wherein the composition is administered 2-7 days after completion of the lymphodepleting chemotherapy regimen. Page 170 of 173 12403779v1Attorney Docket No. 2017428-0684 63. The method of claim 61 or claim 62, wherein the composition is administered 2 days after completion of the lymphodepleting chemotherapy regimen.

64. The method of any one of claims 61-63, wherein the lymphodepleting chemotherapy is or comprises cyclophosphamide and fludarabine.

65. The method of any one of claims 1-64, further comprising: collecting a sample from the subject after administration of the composition and determining the presence of cellular and / or humoral immune response directed towards one or more CD22-specific CAR peptides.

66. The method of claim 65, wherein the method further comprises modulating a subsequent administration of the composition, a post-treatment therapy, and / or one or more additional anti-cancer therapies based on the presence of cellular and / or humoral immune response.

67. The method of any one of claims 1-66, further comprising: collecting a sample from the subject after administration of the composition and determining: CAR T enumeration, CAR T cellular kinetics, CD47 expression, presence of replication competent lentivirus, T cell clonality, immunophenotype, and / or a cytokine profile.

68. The method of claim 67, wherein the method further comprises modulating a subsequent administration of the composition, a post-treatment therapy, and / or one or more additional anti-cancer therapies based on the CAR T enumeration, CAR T cellular kinetics, CD47 expression, presence of replication competent lentivirus, T cell clonality, immunophenotype, and / or cytokine profile.

69. The method of any one of claims 1-68, further comprising: collecting a sample from the subject after administration of the composition and determining the presence of a: hypoimmunogenic T cell-mediated NK response, hypoimmunogenic T cell-mediated T response, hypoimmunogenic T cell-mediated humoral response, and / or humoral immunogenic response.

70. The method of claim 69, wherein the method further comprises modulating a subsequent administration of the composition, a post-treatment therapy, and / or one or more additional anti-cancer therapies based on the Page 171 of 173 12403779v1Attorney Docket No. 2017428-0684 presence of the hypoimmunogenic T cell-mediated NK response, hypoimmunogenic T cell-mediated T response, hypoimmunogenic T cell-mediated humoral response, and / or humoral immunogenic response.

71. An article of manufacture comprising a composition of a cell therapy, or one of a plurality of compositions of a cell therapy, comprising T cells expressing an anti-CD22 chimeric antigen receptor (CAR), and instructions for administering the cell therapy, wherein the instructions specify administering the T cell composition according to the methods of any of claims 1-70.

72. A pharmaceutical composition comprising a population of engineered hypoimmunogenic T cells comprising: (a) reduced expression of beta-2-microglobulin (B2M), Class II Major Histocompatability Complex Transactivator (CIITA), and T cell receptor alpha constant (TRAC) relative to a control or wild-type T cells, (b) increased expression of CD47 encoded by a first exogenous polynucleotide relative to the control or wild-type T cells, and (c) expression of a CD22-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.

73. The pharmaceutical composition of claim 33, wherein the pharmaceutical composition comprises partially edited CAR+ cells, wherein the partially edited CAR+ cells comprise one or more modifications that: (a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and / or (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules, or (b) increase expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not comprise the modification, and express a CD22-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide. Page 172 of 173 12403779v1

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